NCBI

Skip to main content

Skip to navigation

Resources

How To

About NCBI Accesskeys

Sign in to NCBI

PMC
US National Library of Medicine
National Institutes of Health
Search database
Search term

Advanced

Journal list

Help

Journal List

Braz J Microbiol

v.40(1); Jan-Mar 2009

PMC3768498

Braz J Microbiol. 2009 Jan-Mar; 40(1): 111.

Published online 2009 Mar 1. doi: 10.1590/S1517-83822009000100001

PMCID: PMC3768498

Principles and applications of polymerase

chain reaction in medical diagnostic fields: a
review
Marcela Agne Alves Valones,1,* Rafael Lima Guimares,1 Lucas Andr Cavalcanti Brando,1
Paulo Roberto Eleutrio de Souza,2 Alessandra de Albuquerque Tavares Carvalho,1 and Sergio
Crovela1
Author information Article notes Copyright and License information
This article has been cited by other articles in PMC.
Go to:

Abstract
Recent developments in molecular methods have revolutionized the detection and
characterization of microorganisms in a broad range of medical diagnostic fields, including
virology, mycology, parasitology, microbiology and dentistry. Among these methods, Polymerase
Chain Reaction (PCR) has generated great benefits and allowed scientific advancements. PCR is
an excellent technique for the rapid detection of pathogens, including those difficult to culture.
Along with conventional PCR techniques, Real-Time PCR has emerged as a technological
innovation and is playing an ever-increasing role in clinical diagnostics and research laboratories.
Due to its capacity to generate both qualitative and quantitative results, Real-Time PCR is
considered a fast and accurate platform. The aim of the present literature review is to explore the
clinical usefulness and potential of both conventional PCR and Real-Time PCR assays in diverse
medical fields, addressing its main uses and advances.
Keywords: Conventional PCR, Real-Time PCR, Molecular Biology, Methods of molecular
detection
Go to:

INTRODUCTION
The development of molecular biology was one of the greatest achievements in biological
science in the century XX. The discovery of Polymerase Chain Reaction (PCR) brought
enormous benefits and scientific developments such as genome sequencing, gene expressions in
recombinant systems, the study of molecular genetic analyses, including the rapid determination
of both paternity and the diagnosis of infectious disease (73,99). PCR enables the in vitro
synthesis of nucleic acids through which a DNA segment can be specifically replicated in a semiconservative way. It generally exhibits excellent detection limits (19,101).

Recently, a technological innovation of PCR, known as Real-Time PCR, has become

increasingly important in clinical diagnostics and research laboratories due to its capacity for
generating quantitative results. This technique allows accompanying the reaction and
presentation of results in a faster and more accurate fashion than conventional PCR, which only
displays the qualitative results (50,62,73).
The aim of the present study is to outline the principles and applications of conventional PCR
and Real-Time PCR techniques in some medical sciences. It also seeks to evaluate and discuss
the indications, uses and advantages of these techniques, as well as their advances in various
medical areas.
Go to:

PCR TECHNOLOGY CONVENCIONAL PCR

PCR was developed in the 1980s by Kary Mullis, who received the Nobel Prize in 1994 (14).
Since its description, this technology has caused a veritable revolution in biological research,
establishing the agreement of basic biological processes in applied areas involving diagnoses and
genetic improvements for plants and animal (101). PCR enables the synthesis of specific DNA
fragments using a DNA-polymerase enzyme, which takes part in the replication of the cellular
genetic material. This enzyme synthesizes a complementary sequence of DNA, as a small
fragment (primer) is connected to one of the DNA strands in the specific site chosen to start the
synthesis. Primers limit the sequence to be replicated and the result is the amplification of a
particular DNA sequence with billions of copies (66,73).
The development of tools for amplifying DNA segments has generated enormous benefits in
gene analysis as well as the diagnosis of many genetic diseases and the detection of bacterial,
viral and fungal pathogens (4,72,73,99). Another useful PCR application is the cloning of a
particular DNA fragment, which allows the study of gene expression and has considerable
potential in forensic medicine (94).
Go to:

REAL-TIME PCR
The possibility of Real-Time PCR monitoring has revolutionized the quantification process of
DNA and RNA fragments. Real-Time PCR allows the precise quantification of these nucleic
acids with greater reproducibility. This technique provides a sensitive method for the accurate
quantification of individual species, which could be very relevant to the diagnosis of pathogens
and genetic diseases. Advantages of Real-Time PCR include the ease of quantification, greater
sensitivity, reproducibility and precision, rapid analysis, better control of quality in the process
and a lower risk of contamination (62,73).
Real-Time PCR requires a thermocycler with an optical system to capture fluorescence and a
computer with software capable of capturing the data and performing the final analysis of the

reaction. The programs available from diverse manufactures exhibit differences regarding sample
capacity, method of excitation and total sensitivity. There are also differences between regarding
the data processing. The emission of fluorescence generates a signal that increases in direct
proportion with the amount of PCR products. Fluorescence values are recorded during each cycle
and represent the amount of amplified product. The fluorescent composites used are SYBR
Green and TaqMan (50,73).

Real-Time PCR Technology

Forms of detection
The fluorescence signals are proportional to the amount of PCR product generated by the
fluorescent dyes, which are specific to double-stranded DNA (dsDNA), or by sequence-specific
oligonucleotide probes.

SYBER Green I dye

SYBR Green I is the most used dsDNA binding-specific dye in real-time PCR. Its fluorescence is
undetectable when not bound to dsDNA. Its binding affinity to DNA is 100 times greater than
that of ethidium bromide, which is the most often used dsDNA binder in conventional PCR
(64,113). The disadvantage to SYBR Green I is that it binds to any dsDNA, such as non-specific
amplification products and primer dimers. Amplified non-specific products affect the efficiency
of the amplification of specific products. Thus, analysis should be optimized in such a way that
non-specific amplification does not occur. Melting curve analysis after the PCR reaction is a
good practice for controlling the formation of dimer primers. Fluorescence is measured as a
function of temperature, gradually diminishing with the increase in temperature of the amplified
product (74). However, upon reaching the temperature at which the double-stranded DNA
separates, the stain detaches and fluorescence drops off abruptly (84). Once optimized, detection
by SYBR Green I is highly sensitive to the identification of a single molecular target in the
reaction mixture. The greatest advantage is that it can be used with various pairs of different
primers, making it less expensive than a probe.

Minor groove DNA binder probes (MGB)

MGB probes consist of oligonucleotides from 14 to 15 pb in length that carry a fluorescent dye
in terminal 5 as well as a non-fluorescent quencher and MBG in terminal 3, which specifically
hybridizes with a target sequence. MGB is released from a probe that binds to the minor groove
of the dsDNA (consisting of part of the MGB probe and complementary target sequence by
which it is hybridized) related to the nucleotide sequence. The MGB increases binding stability
to the amplification probe (51).

Hybridization probes
Oligonucleotide probes marked with fluorophores are used for the detection of specific
sequences (16,115). The amount of the fluorescence may be related to the amount of PCR
product through the product-dependent reduction of a quencher fluorophore and a reporter or

through an increase in the fluorescent resonance energy transfer (FRET) from a donor
fluorophore to a receptor. The donor probes are marked in the 3 terminal portion with a reporter
fluorophore (often 6-carboxifluorescein, FAM) and the acceptor probes are marked in the 5
terminal portion with an acceptor fluorophore (cyan dyes Cy3, Cy5; 6-carboxy-4,7,2,7tetrachlorofluorescein, TET; 6-carboxy-N, N, N, N-Tetramethylrhodamine, TAMRA; 6carboxirhodamine X, ROX). Only the donor fluorophore is excited in such a way that no
fluorescent acceptor is detected in the free-floating probes. During the annealing phase of the
primer, the probes hybridize adjacently to the single-stranded DNA (ssDNA) and the excitation
energy is transferred from the donor to the acceptor. Four oligonucleotides are used in this
format: two primers and two probes. The amount of fluorescence is proportional to the amount of
target DNA generated during the PCR process (10).

TaqMan Probes
The first fluorescent probe developed for real-time PCR was 5nuclease, which is commonly
referred to by the name TaqMan. A TaqMan probe is a short oligonucleotide (DNA) that contains
a 5terminal reporter fluorophore similar to fluorescein and a 3terminal quencher. Intact probes
do not emit fluorescence because they are bound (quenched). Two events must occur to generate
a fluorescent signal. First, the probe must bind to a complementary DNA strand at 60C. Second,
at this temperature, Taq polymerase, the same enzyme used for the PCR, must cleave the 5
terminal TaqMan probe (5nuclease activity), separating the fluorescent dye from a quenching
dye. The quencher is released from the fluorophore, which now fluoresces after excitation
(28,33).

Molecular beacons
Tyagi, Kramer (106) first evaluated molecular beacons combine an oligonucleotide capable of
forming a stem-loop structure with the quencher-reporter pair. Specifically, an oligonucleotide
probe with a binding domain to the antisense target flanked by two short arms of complementary
sequences is marked in one terminal with the reporter dye and in the opposite terminal with the
quencher dye. In the absence of the target, the short arms anneal to form a hairpin structure
(stemloop), forcing the flourophore toward the quencher. In this conformation, the molecular
beacon is dark. Through hybridization with the target sequence, the hairpin structure opens,
separating the fluorophore and quencher and resulting in the restoration of fluorescence
(shining state) (107). The transition between the dark and shining state of the molecular beacon
allows the differentiation between bound and unbound probes (49,106,108).

Sunrise primers
SunriseTM Probes (Oncor, Inc.) (70) function in a similar manner to TaqMan probes. They are
also doubly marked with a flourochrome in the 5terminal portion. The 3 region of the probe is
the specific target and the 5 region is the complementary target, such that when not extended
(not incorporated in the amplicon), it forms a hairpin structure that holds the quencher and
reporter marker together. When the probe is extended and integrated within a dsDNA molecule,
the quencher and reporter are kept apart by a recently copied complementary strand. Like
conventional TaqMan, Sunrsire primers require a new probe for each amplification (109).

In the first phase, the Sunrise primer is extended with the forward primer. This extended product
serves as the template for the reverse primer in the second phase. In the end, polymerase opens
the hairpin structure and a double-stranded PCR product is formed, in which the reporter and
quencher are separated (70).

Scorpion primers
Scorpion primers are structurally and functionally related to molecular beacons, but serve as
primers in PCR. Two different formats are possible: stem-loop and duplex. In both cases, the
marking mechanism is intra-molecular. The basic elements of scorpions are: (i) a PCR primer;
(ii) a PCR stopper to prevent the cross-reaction of a probe; (iii) a sequence-specific probe; and
(iv) a fluorescence detection system containing a minimum of one flourochrome and one
quencher. After PCR amplification of the scorpion primer, the resulting amplicon contains a
sequence that is complementary to the probe, which is restituted a single strand during the
denaturation stage of each PCR cycle. With cooling, the probe is free to bind to this
complementary sequence, producing increased fluorescence. Thus, the quencher is not increased
in the proximity of the fluorophore (110).
Go to:

PRINCIPLES AND APPLICATIONS IN VIROLOGY

Recent advances in molecular biology have made possible the detection and characterization of
viral nucleic acids. Methods such as PCR enable the amplification of specific regions of interest.
Technological improvements in the detection systems of gene sequences provide a complete viral
characterization, determining the subtype, genotype, variation, mutation and standards of
genotypic resistance of these viruses (56,71). The recent development of Real-Time PCR has
facilitated the detection and amplification of PCR products. This method is useful in quantifying
a larger range of sequences of viral nucleic acids than most quantitative methods. Moreover, the
qualitative detection also is possible. Quantification and qualification are carried out
automatically. Examples of the detection and quantification of specific viral regions have been
published and this field of study is growing very quickly (39,72).
The implementation of molecular methods has resulted in progress regarding the diagnosis of
many viruses and the monitoring of antiviral therapy, especially HIV-1 (Human
Immunodeficiency Virus type 1) (92), HBV (Hepatitis B Virus) (79) and HCV (Human
Cytomegalovirus) (23). It has also led to the development of amplification assays on nearly all
human viruses, including those that are more easily be cultivated, such as HSV-1 and HSV-2
(Herpes Simple Virus type 1 and 2) (25).
The newer molecular methods are advantageous, mainly in cases for which the viral culture
routine is not available. The introduction of molecular biology in clinical diagnosis is important
to reducing the use of viral culture techniques. The implementation of automatic extraction and
detection, combined with an extensive quality control program, should convince the clinical
community that molecular diagnosis is important in clinical virology (46,120). The ability to
exclude viral infections can help avoid unnecessary therapies, such as powerful antibiotics and

antiviral medicines, as well as reduce costs incurred on the part of patients. Thus, these
techniques are important to establishing the best therapeutic protocol (56,72).
Real-Time PCR is extremely useful in the study of viruses that cause infectious diseases. The
majority of published assays show an increase in the frequency of viral detection. This is
therefore an attractive technology for many virological fields (56). It is valued for its quickness
in the detection of viral variants and the syndromes caused by these viruses (27). The method
contributes to epidemiological studies due to its capacity to quantify nucleic acids in a single
reaction (46,120). New chemicals have allowed a better discrimination of multiple viral
genotypes within a single reaction (44) and have provided an alternative viral detection method
based on morbidity and mortality assays.
For many years, the diagnosis of viral infections has been hampered by the high costs, laboratory
time and qualified personnel required in the cell culture process. An additional negative factor is
the low sensitivity and slow development of many viruses in artificial mediums. PCR technology
facilitates and improves detection, thereby facilitating the diagnosis of a certain number of these
viruses (100).
Go to:

PRINCIPLES AND APPLICATIONS IN MYCOLOGY

AND PARASITOLOGY
The ability to accurately identify microorganisms is fundamental to all aspects of fungal
epidemiology and diagnosis. In phytopathology, the early identification of disease-causing agents
is essential to the recognition of pathogens (60). In the last ten years, advancements have been
made in the molecular diagnosis of fungi through PCR technology. Unlike conventional
methods, samples can be tested directly through PCR and isolated without the need for cultures.
The technique is fast and highly specific. It can be used to detect trace amounts of fungal DNA
from environment samples before symptoms occur. It therefore allows the implementation of
early disease control methods. PCR can be performed routinely and does not require specialized
skill to interpret the results. The technology can also offer more accurate quantitative data,
providing additional information necessary for decision making and the assessment of how
effective fungal agents are in biological control. Since its introduction in the mid 1980s, PCR has
become the cornerstone of DNA technology and has cleared the path for the creation of
innumerous associated technologies. It is remarkable for its ability to detect amounts of DNA
amplified from one or few original sequences. Conventional PCR is not quantitative, but rather
qualitative. It has been used to detect, monitor and identify fungi from an entire set of
environmental samples and is the core of molecular fungal diagnostics (4).
Fluorescent PCR in situ utilizes fluorescently marked primers or probes to detect and locate
fungi in fixed environmental samples following semi-permeabilization (102). The fluorescence
of the primers or probes is detected using a confocal microscope. This technique allows the
direct detection of the organism in the sample. It also shows the spatial distribution, interactions
with the host and other organisms. Bago, Piche, Simon (5) used in situ PCR to detect and locate

infections caused by Arbuscular Mycorrhiza fungi. The scope of PCR is infinite (5). It can be
used to investigate either a single species or entire communities (22,35,80).
Pneumocystis jiroveci (a fungus previously denominated Pneumocystis carinii) can cause severe
pneumonia in patients infected with HIV or otherwise immunosuppressed, but its detection is
restricted to the microscopy of specimens in the respiratory tract. Microscopy for the detection of
P. jiroveci generally involves the use of stains. Immunofluorescence is more sensitive than these
stains, but is more expensive and requires specialized facilities. PCR is more sensitive, especially
in patients not infected with HIV, and can therefore be of considerable usefulness (36). PCR
specificity is limited, but as this microorganism is an omnipresent commensal, it can be detected
through PCR in the absence of the disease (37).
Another example of the use of PCR technology in mycology is in the detection of infection from
Aspergillus ssp. in patients with neutropenia. This disease is notoriously difficult to diagnose due
to the poor sensitivity of the culture method and the difficulty of finding histopathological
specimens in individuals with low platelet counts. Early treatment is essential to achieving the
best results. PCR can reduce the time required for the specific diagnosis (111). Real-Time PCR
has been successfully used to quantify the number of pathogens (7,13,21,112), thereby assisting
in decisions regarding how to treat fungal diseases and assess the effects of fungi (57).
Parasitological diagnostics can be assisted by molecular methods. Many parasites are not
cultivable in laboratory and diagnosis principally relies on serology and relatively less sensitive
microscopy. Microscopy remains a support to the diagnosis of malaria, but due to its greater
sensitivity, PCR can diagnose this illness in even in difficult situations. Plasmodium species can
also be detected in different infections, which can hinder microscopic discernment (99)
Go to:

PRINCIPLES AND APPLICATIONS IN

MICROBIOLOGY
Conventional PCR has been used for over a decade in clinical microbiology laboratory research
for the identification of microbial pathogens (114). However, for a number of reasons, this
technique has been restricted to the detection of microorganisms that either have slow growth or
cannot be cultivated. Most tests based on conventional PCR involve multiple steps and,
therefore, require careful expertise. These assays often require both time and culture-based
methods, thereby increasing the costs. Conventional PCR also involves an open-reaction system,
which is more susceptible to contamination from foreign amplified DNA. Conventional PCR
assays have been developed for Bacillus anthracis, the Anthrax agent (9) and Variola Major (26),
but clinical validation of these assays is limited due to the unavailability of the human specimen.
Real-time PCR has important, immediate implications to diagnostic tests in the clinical
microbiology laboratory. The enhanced sensitivity, ease of use and quickness of this technology
make it an attractive alternative for detecting microorganisms in humans (18).

Bacteriology

Anaerobic bacteria are involved in a broad range of infections that are commonly associated to
considerable morbidity and mortality rates (98,116). Although different types of these bacteria
are frequently found in diverse infections, evidence suggests that a number of these clinically
important pathogens are as yet poorly characterized due to the inadequacy of conventional
anaerobic bacteriological methods and phenotype tests. According to recent studies, 50 to 75
percent of anaerobic bacteria are satisfactorily characterized and 27 percent of laboratories
indicated that they have never identified such bacteria (2). This occurs mainly because
conventional identification is complicated, expensive and time-consuming. It is also not reliable,
as it is based on antiquated taxonomy (97).
Molecular detection methods are a powerful means of identifying these pathogens in the study of
the parasite-host relationship, clarifying the taxonomic positions of known pathogens. Thus,
there is a growing trust in genotyping for microbial characterization. Genotypes are more
specific and more easily quantified and standardized between different organisms than traditional
phenotype markers. In the last ten years, a number of advances have been made in molecular
bacterial diagnostics, including the greatest discovery: PCR technology (97,116). Real-time PCR
has recently emerged and has been used in the detection and quantification of anaerobic bacteria.
This provides users with the ability to amplify DNA as well as detect and confirm the specific
sequences of microorganisms. Each cycle in real time also provides greater sensitivity.
PCR for the detection of Lactobacillus, Gadnerella vaginallis and Mycoplasmas hominis, genital
tract bacteria was used (119). In another study, Aliyu et al. (1) reported the importance of
Fusobacterium necrophorum as a cause of acute pharyngitis. There are a considerable number of
studies using real-time PCR in the characterization of bacterial communities in the human
intestine (6,40,58).
The diagnosis of infections due to specific bacteria has greatly benefited from molecular
detection. Many of these bacteria are public health concerns, such as Micobacterium
tuberculosis, Chlamydia Trachomatis, Neisseria gonorrheae and Bordetella pertussis. Tests
based on molecular methods have the advantage of avoiding days or weeks of delay and allow
early recognition and treatment. Commercial assays are available for M. tuberculosis,
Micobacterium avium complex, C. trachomatis and N. gonorrheae (116).
Due to the increased sensitivity, the use of molecular detection methods for sexually transmitted
bacteria has led to an increase in the proportion of cases confirmed in laboratory of the diseases
such bacteria cause. Traditional sexual health exams require the use of a speculum in women and
a urethral swab in men. These exams require special equipment and can cause both
embarrassment and discomfort. Molecular detection is a noninvasive method, which increases
trust and reduces discomfort. Molecular testing also encompasses a range of genital pathogens,
such as C.Trachomatis, N. gonorrheae, etc. (100).
Go to:

PRINCIPLES AND APPLICATIONS IN DENTISTRY

The recognition of the universality of the genetic code in living organisms has been essential to
the development and application of genetic technologies. A number of methods have been
employed in dentistry to better understand and diagnose infectious agents that lead to
maxillofacial infections, thereby obtaining an evaluation of risk patients might have to caries,
periodontal disease and other oral conditions (47). These methods include cultures, microscopy,
immunofluorescence assays and DNA probes. More recently, polymerase chain reaction has been
introduced. Many different types of clinical samples have been used for PCR analyses, including
blood, sweat, semen, strands of hair and saliva. Saliva is a potential source of genetic material for
diagnostic tests in oral and systemic diseases. With the recent interest in associations between
systemic disease and the mouth as well as the successful use of saliva in molecular diagnostics,
saliva sampling may soon become part of the routine dental exam. Furthermore, as a medium
where biologically active proteins and exogenous substances are found, saliva is also a source of
patient DNA (82,88,103) as well as cariogenic and periodontopathgenic microorganisms
(32,42,48,76).
The publication of the PCR technique was a revolutionary watershed for medicine and science. It
has recently become a standard diagnostic and research tool in dentistry. The literature reports
the uses of PCR for the study of factors involved in periodontal disease, dental caries, endodontic
infections and oral cancer (90).

Periodontal disease
Diverse methods have been reported for the rapid detection of periodontal pathogens, such as
immunological and immunoenzymatic assays, protein electrophoresis and DNADNA
hybridization. However, these methods exhibit different limitations that can lead to false positive
results as well as cross-reactions (3,17).
PCR is an excellent tool for directly identifying periodontal pathogens in subgingival samples.
Due to its sensitivity and specificity, it is also a rapid, efficient method for detecting, identifying
and differentiating organisms, but appropriate standardization is necessary (24). Diverse
molecular means are often used to identify periodontal pathogens, but PCR is considered the
easiest and fastest method in clinical samples (83). PCR may soon become the ideal detection
method for periodontal pathogens due to its greater ease of use in comparison to cultures
associated with biochemical identification tests. It also demonstrates excellent detection limits
with few cross-reactions under ideal conditions (19).
In researching the possible involvement of human viruses in periodontal disease, Parra, Slots
(77) determined the prevalence of Human Cytomegalovirus (HCMV), Epstein-Barr Virus Type I
and II (EBV 1 and 2), Herpes Simples Virus (HSV), Human Papillomavirus (HPV) and Human
Immunodeficiency Virus (HIV) in the crevicular fluid of individuals with various forms of
periodontal disease. Viral identification was carried out using the PCR technique. The study
provided evidence of human viruses in the crevicular fluid of the most advanced adult
periodontal lesions. Ssygun et al. (91) confirmed the frequent presence of HCMV in crevicular
samples of chronic periodontal lesions and suggested a strong relation between HCMV and
EBV-1 in subgingival areas with deep furrows accompanied by a loss of insertion.

In studying the expression of virulent factors of Porphyromonas gingivalis in periodontitis,

Shelburne et al. (93) modified methods of quantitative measurements and gene activation, and
discovered (real-time) quantitative reverse transcription of PCR (QRT-PCR) to examine gene
expression in vivo in the oral cavity of the anaerobe P. gingivalis. The authors described these
initial results with QRT-PCR using a selection of virulent factors. The effectiveness of the PCR
method in the detection of P. gingivalis from saliva samples was compared to bacterial cultures
performed by Matto et al. (59), demonstrating that PCR detected the bacteria in saliva three time
more frequently than the culture method.
The PCR was also used by Sakamoto et al. (89) for the detection and identification of
Treponema socranskii associated to periodontal disease. This pathogen is related to an
inflammation in the gingiva in both children and adults. T. socranskii was detected more
frequently in subgingival plaque samples than in saliva samples.
Studies have detected Actinobacillus actinomycetemcomitans using PCR in the most distinct
patterns in a population with (70%) and without (19%) periodontal disease. These studies have
also shown the sensitivity and specificity of PCR in comparison to traditional culture methods
(65,104). Okada et al. (75) used PCR to detect the presence of A. actinomycetemcomitans and P.
gingivalis in dental plaque samples from children using their toothbrushes. The research
indicated that such bacteria are rarely present in the oral cavity of healthy children. In 2001, the
researchers also detected the presence of P. Intermdia, nigrescens, B. forythus, T.denticola and
C. rectus in periodontal disease.
Real-Time PCR offers a sensitive, efficient, faithful approximation for quantification. Using the
TaqMan systems, Lyons et al. (54) were able to determine both the amount of P. gingivalis and
total number of bacteria present in bacterial plaque without the use of cultures. Furthermore, this
allowed the authors to determine the percentage of P. gingivalis in a complex sample.
Sakamoto et al. (88) compared conventional PCR, real-time and the culture method in the
detection and quantification of periodontopathogenic bacteria, including A.
actinomycetemcomitans, B. forsythus, P. gingivalis, T. denticola and T. socranskii in both saliva
and subgingival plaque. There was satisfactory agreement between the results from conventional
and real-time PCR for all the saliva samples. The use of real-time PCR optimally simplified the
process and was able to determine the amount of periodontopathogenic bacteria within one hour.
These bacteria were more frequently detected in the saliva than the subgingival plaque. The
study suggests the saliva is as good, or better, than subgingival plaque for the detection and
quantification of periodontal bacteria in the oral cavity.
Therefore, PCR has revolutionized understanding regarding periodontal pathogens. These studies
not only permit the diagnosis of known pathogens, but also contribute toward the identification
of new pathogens involved in periodontal disease (47). PCR is expected to be successful in the
microbiological diagnoses and periodontal disease (89,90).

Dental Caries

The group mutans, which encompasses cariogenic bacteria, has seven species: Streptococcus
cricettus, S. ratti, S. mutans, S. sobrinus, S. downei, S. ferus and S. macacae (69,108), of which
S.mutans and S. sobrinus are more frequently isolated from the human oral cavity (38,108,117).
For epidemiological studies on caries, a rapid, sensitive, simple method is needed for the
identification and differentiation of these microorganisms. A number of methods have been used
to identify these isolated species in the oral cavity, such as biochemical, immunological and
genetic methods (41). Different techniques are used in conjunction in order to achieve reliability
in species identification. This requires time and considerable ability, and the results are often
unsatisfactory. Species characterization based on DNA testing is currently widely accepted, as
phenotyping is a reflection of gene expression (105). As mentioned above, PCR was recently
introduced to solve these problems. Tests related to genotyping are currently being used due to
the possibility of its being more sensitive and specific (118).
PCR appears to be convenient for studying the epidemiology of disease in isolated individuals. It
may prove useful in the identification of species associated with dental caries and their location
in the ecological niches, thereby helping to clarify the progression of the carious process (82).
PCR has the potential to replace conventional identification methods, such as biochemical and
immunological tests (43) The discriminative power of PCR in the differentiation of S. mutans
and S. sobrinus serotypes and lineages was investigated by Saarela et al. (87), who found that
PCR exhibited good results in differentiating S. mutans lineages and the technique is appropriate
for epidemiological studies on this bacterium.
Rupf et al. (86) presented a competitive PCR method for the specific quantitative determination
of S. mutans. This method allowed a quick, exact determination of unknown quantities of this
bacterium and provided an efficient means for evaluating the risk of caries in patients as well as
monitoring the efficiency of preventative and therapeutic measures.

Endodontic Infections
With the development of molecular methods based on the detection of specific genomic regions,
it became possible to identify microbial species in infected root canals that had never been seen
by means of the conventional culture procedure. Examples of bacterial species that were only
detected in canals through molecular methods and that are currently considered important
endodontic pathogens include Treponema denticola, Dialister pneumosintes, Filifactor alocis,
Tannerella forsythia, Treponema malthophilum, Treponema socranskii and Prevotella tannerae
(96). Based on current studies, molecular methods can be used to characterize the micro-flora
associated to endodontic infections (47). Using PCR, Bogen, Slots (12) sought to determine the
prevalence of P. endodontalis, P. gingivalis, P. intermdia and P. nigrescens in 20 with refractory
periapical lesions. The authors concluded that bacteria that produce black pigment do not appear
to constitute the majority of microorganisms in these cases. The relatively lower occurrence of
these bacteria may help explain the relative stability and chronic nature of this condition.
Molecular methods such as PCR are more sensitive and specific than the conventional culture
method, which is based on identification through the phenotypic characteristics of each microbial
species (97). Molecular methods, however, are based on the detection of specific genomic
sequences for each microbial species and can even discriminate clones within a single species

(85). Although studies have determined that bacteria are the greatest etiological agent in pulp and
periradicular disease, fungi have also been associated to root canal infection. Baumgartner et al.
(8) used PCR to assess the content of infected root canals as well as cellulite aspirations and
abscesses of endodontic origin for the presence of Candida albicans. The results indicate that
PCR is an extremely sensitive molecular method and can be used to identify C. albicans directly
in samples of endodontic infection.
PCR has been widely used to identify microbial species that are difficult or impossible to
cultivate, as well as colonies within a species that exhibit a different phenotypic behavior and are
therefore difficult to identify in culture procedures. PCR has a greater detection pattern than
traditional microbiological identification methods and exhibits greater specificity under
optimized conditions. Thus, the use of identification methods based on the knowledge of
molecular biology has revolutionized medical microbiology and is broadening the horizons with
regard to the actual profile of endodontic infection (85,95).

Oral Cancer
One of the uses of PCR in dentistry is the detection of markers in the diagnosis and prognosis of
some types of oral cancer (45,52,81). For this purpose, PCR is a fast, easy method with a
relatively low cost (45). Diagnoses, prognoses and treatment can be improved through the study
and use of genetic markers identified by means of immunohistochemistry, PCR and other
molecular biology procedures (67).
Squamous cell carcinoma of the oral cavity is generally accompanied by other types of
aerodigestive tract carcinomas, such as oropharyngeal and esophageal carcinoma. Streptococcus
anginosus is a bacterium that may be isolated in different parts of the body and has been isolated
in squamous cell carcinoma of the head and neck. Through real-time PCR, S. anginosus can be
detected with greater sensitivity and specific approximation in squamous cell carcinomas of the
oral cavity (63).
Pre-malignant lesions of the head and neck have been studied extensively through genetic
alterations. A genetic progression model has been established based on histological alterations
that occur in the interior of the pre-malignant epithelium (15). HPV (Human Papillomavirus) has
been employed in tumor progression in humans based on data from patients with cancer (29).
Determining the moment of viral infections in these premalignant lesions could clarify the role of
HPV in carcinogenesis and help guide future strategies for the prevention and early detection of
squamous cell carcinoma of the head and neck. For such, a large number of studies have been
conducted to detect the presence of HPV in head and neck epithelia through the use of a variety
of laboratorial methods. Different techniques have been employed, including PCR, in situ
hibridization, etc. Real-time PCR minimizes the risk of contamination, thereby becoming the
ideal assay for HPV DNA detection (31).
The quantification of the number of genetic copies through real-time PCR using either DNA or
RNA in studies on gene expression has been reported in the literature for human tumors,
including breast cancer (11,53,61), follicular lymphoma (30), stomach cancer (68), prostate
cancer (34) and Ewings sarcoma (78). In carcinoma of the head and neck, this new technology

has been mainly applied for the detection of Epstein-Barr virus in nasopharynx cancer (55) and
squamous cell carcinoma in lymph nodes (20).
Go to:

CONCLUSIONS
Based on the items research and described above, the following may be concluded:

Methods associated to molecular biology have made excellent progress, with clear
usefulness in diverse fields of medical science. The discovery of Polymerase Chain
Reaction (PCR) introduced an technological advancement that is relevant for the
detection of microorganisms, increasing the sensitivity, precision and accuracy of the
diagnosis;

In virology, the molecular detection and characterization of viral nucleic acids makes a
complete viral characterization possible, thereby providing greater knowledge regarding
the behavior of the virus and selected infectious processes. In the diagnosis of viruses,
this enhances therapeutic treatment as well as clinical and epidemiological virus studies,
thereby avoiding unnecessary treatments and reducing overall costs for patients;

In mycology and parasitology, PCR technology favors the early identification of

microorganisms, thereby enhancing epidemiological studies as well as the diagnosis of
fungi and parasites, which is essential to the recognition of pathogens. Molecular tests
assist in the decision-making process regarding fungal and parasitological diseases by
assessing the effects of these microorganisms;

In microbiology, PCR permits important, immediate observations for diagnostic tests in

the detection of microorganisms. Genotyping allows the study of bacteria such as
Mycobacterium tuberculosis. This is of tremendous worth for public health, favoring the
early recognition and optimized treatment;

In dentistry, molecular methods enhance knowledge regarding the diagnosis of

infectious agents that lead to maxillofacial infections, thereby favoring the assessment of
patients at risk for conditions such as caries, periodontal disease, endodontic infections
and oral cancer. The publication of the PCR technique was a revolutionary watershed for
medicine and science. It has become a standard diagnostic and research tool in dentistry,
permitting the early diagnosis of the above-mentioned diseases.

Go to:

RESUMO
Princpios e aplicaes da reao em cadeia de Polimerase em reas mdicas diagnsticas:
uma reviso

O advento dos mtodos moleculares tem, nos ltimos anos, revolucionado a deteco e
caracterizao dos microorganismos em diversas reas mdicas diagnsticas, tais como
virologia, micologia, parasitologia, microbiologia e odontologia. Dentre as tcnicas baseadas em
biologia molecular, a PCR (Polymerase Chain Reaction) trouxe enormes benefcios e
desenvolvimentos cientficos, se mostrando como um excelente caminho para a rpida deteco
de patgenos, at mesmo aqueles de difcil cultivo. Derivada da PCR convencional, a PCR em
Tempo Real se mostra como uma inovao tecnolgica e vem conquistando espao nos
diagnsticos clnicos e nos laboratrios de pesquisa por apresentar a capacidade de gerar, alm
de resultados qualitativos, resultados quantitativos, se mostrando de forma mais rpida e precisa.
Este trabalho de reviso tem por objetivo explorar a utilidade clnica da tcnica de PCR
convencional e em Tempo real nas diversas reas mdicas supracitadas, abrangendo seus
principais usos e avanos, direcionando para o cotidiano profissional.
Palavras-chave: PCR convencional; PCR em Tempo Real; Biologia molecular; mtodos de
deteco molecular.
Go to:

REFERENCES
1. Aliyu S.H., Marriot R.K., Curran M.D., Parmar S., Bentley N., Brown N.M., Brazier J.S.,
Ludlam H. Real-time PCR investigation into the importance of Fusobacterium necrophorum as a
cause of acute pharyngitis in general practice. J. Med. Microbiol. 2004;53((Pt 10)):10291035.
[PubMed]
2. Andrews J.M., Brown D., Wise R. A survey of antimicrobial susceptibility testing in the
United Kingdom. J. Antimicrob. Chemother. 1996;37(1):187188. [PubMed]
3. Ashimoto A., Chen C., Bakker I., Slots J. Polymerase chain reaction detection of 8 putative
periodontal pathogens in subgingival plaque of gingivitis and advanced periodontitis lesions.
Oral Microbiol. Immunol. 1996;11(4):266273. [PubMed]
4. Atkins S.D., Clark I.M. Fungal molecular diagnostics: a mini review. J. Appl. Genet.
2004;45(1):315. [PubMed]
5. Bago B., Piche Y., Simon L. Fluorescently-primed in situ PCR in arbuscular mycorrhizas.
Mycol. Res. 1998;102(12):15401544.
6. Bartosch S., Fite A., Macfarlane G.T., Mcmurdo M.E. Characterization of bacterial
communities in feces from healthy elderly volunteers and hospitalized elderly patients by using
real-time PCR and effects of antibiotic treatment on the fecal microbiota. Appl. Environ.
Microbiol. 2004;70(6):35753581. [PMC free article] [PubMed]
7. Bates J.A., Taylor E.J.A., Kenyon D.M., Thomas J.E. The application of real-time PCR to the
identification, detection and quantification of Pyrenophora species in barley seed. Mol. Plant.
Pathol. 2001;2(1):4957. [PubMed]
8. Baumgartner J.C., Watts C.M., Xia T. Occurrence of Candida albicans in infections of
endodontic origin. J. Endod. 2000;26(12):6958. [PubMed]
9. Bell C.A., Uhl J.R., Hadfield T.L., et al. Detection of Bacillus anthracis DNA by LightCycler
PCR. J. Clin. Microbiol. 2002;40(8):28972002. [PMC free article] [PubMed]
10. Bernard P.S., Wittwer C.T. Homogenous amplification and variant detection by fluorescent
hybridization probes. Clin. Chem. 2000;46(2):1478. [PubMed]

11. Biche I., Olivi M., Champme M.H., Vidaud D., Lidereau R., Vidaud M. Novel approach to
quantitative polymerase chain reaction using real-time detection: application to the detection of
gene amplification in breast cancer. Int. J. Cancer. 1998;78(5):661. [PubMed]
12. Bogen G., Slots J. Black-pigmented anaerobic rods in closed periapical lesions. Int. Endod. J.
1999;32(3):204210. [PubMed]
13. Bohm J., Hahn A., Schubert R., Bahnweg G., Adler N., Nechwatal J., Oehlann R., Osswald
W. Real-time quantitative PCR: DNA determination in isolated spores of the mycorrhizal fungus
Glomus mosseae and monitoring of Phytophora infestans and Phytophora citricola in their
respective host plants. J. Phytopathol. 1999;147(7)(8):409416.
14. Bruce A., Bray D., Johnson A., Lewis J., Rass M., Roberts K., Walter P. Fundamentos da
biologia celular: uma introduo biologia molecular da clula. Porto Alegre: Artes Mdicas
Sul; 1999.
15. Califano J., Van Der Riet P., Westra W., Nawroz H., Clayman G., Piantadose S., Corio R.,
Lee D., Greenberg B., Koch W., Sidransky D. Genetic progression model for head and neck
cancer: implications for field cancerization. Cancer Res. 1996;56(11):24882492. [PubMed]
16. Cardullo R.A., Agrawal S., Flores C., Zamecnik P.C., Wolf D.E. Detection of nucleic acid
hybridization by nonradiative fluorescence resonance energy transfer. Proc. Natl. Acad. Sci.
USA. 1988;85(23):87908794. [PMC free article] [PubMed]
17. Chen C., Slots J. Microbiological tests for Actinobacillus actinomycetemcomitans and
Porphyromonas gingivalis. Periodontol. 2000. 1999;20:5364. [PubMed]
18. Cockerill F.R. Application of rapid-cycle Real-Time polymerase chain reaction for diagnostic
testing in the clinical microbiology laboratory. Arch. Pathol. Lab. Med. 2003;127(9):11121120.
[PubMed]
19. Cortelli S.C., Jorge A.O.C., Querido S.M.R., Cortelli J.R. PCR e cultura na deteco
subgengival de Actinobacillus actinomycetemcomitans: estudo comparativo. Cien. Odontol.
Bras. 2003;6(2):5864.
20. Cortesina G., Martone T., Galeazzi E., Olivero M., De Stefani A., Bussi M., Valente G.,
Comoglio P.M., Di Renzo M.F. Staging of head and neck squamous cell carcinoma using the
MET oncogene product as marker of tumor cells in lymph node metastases. Int. J. Cancer.
2000;89(3):286292. [PubMed]
21. Cullen D.W., Lees A.K., Toth I.K., Duncan J.M. Conventional PCR and real-time PCR
detection of Helminthosporium solani in soil and potato tubers. Eur. J. Plant. Pathol.
2001;107:387398.
22. Dahllof I. Molecular community analysis of microbial diversity. Curr. Opin. Biotechnol.
2002;13(3):213217. [PubMed]
23. Damen M., Cuypers H.T., Zaaijer H.L., Reesink H.W., Schaasberg W.P., Gerlich W.H.,
Niesters H.G., Lelie P.N. International collaborative study on the second EUROHEP HCV-RNA
reference panel. J. Virol. Methods. 1996;58(1)(2):175185. [PubMed]
24. Doungudomdacha S., Rawlison A., Douglas W.I. Enumeration of Porphyromonas gingivalis,
Prevotella intermedia and Actinobacillus actinomycetemcomitans in subgingival plaque samples
by a quantitative-competitive PCR method. J. Med. Microbiol. 2000;49(10):861874. [PubMed]
25. Espy M.J., Uhl J.R., Mitchell P.S., Thorvlson J.N., Svien K.A., Wold A.D., Smith T.F.
Diagnosis of herpes simplex virus infections in the clinical laboratory by LightCycler PCR. J.
Microbiol. 2000;38(2):795799. [PMC free article] [PubMed]

26. Espy M.J., Cockerill F.R., Meyer R.F., Bowen M.D., Poland G.A., Hadfield T.L., Smith T.F.
Detection of smallpox virus DNA by LigthtCycler PCR. J. Clin. Microbiol. 2002;40(6):1985
1988. [PMC free article] [PubMed]
27. Furuta Y., Ohtani F., Sawa H., Fukuda S., Inuyama Y. Quantitation of varicella-zoster virus
DNA in patients with ramsay hunt syndrome and zoster sine herpete. J. Clin. Microbiol.
2001;39(8):28562859. [PMC free article] [PubMed]
28. Gibson U.E., Heid C.A., Williams P.M. A novel method for real time quantitative RT-PCR.
Genome Res. 1996;6(10):9951001. [PubMed]
29. Gillison M.L., Koch W.M., Capone R.B., Spafford M., Westra W.H., Wu L., Zahurak M.L.,
Daniel R.W., Viglione M., Symer D.E., Shah K.V., Sidransky D. Evidence for a causal PCR in
medical diagnostic fields association between human papillomavirus and a subset of head and
neck cancers. J. Natl. Cancer Inst. 2000;92(9):709720. [PubMed]
30. Goff L.K., Neat M.J., Crawley C.R., Jones L., Jones E., Lister T.A., Gupta R.K. The use of
real-time quantitative polymerase chain reaction and comparative genomic hybridization to
identify amplification of the REL gene in follicular lymphoma. Br. J. Haematol.
2000;111(2):618625. [PubMed]
31. Ha P.K., Pai S.I., Westra W.H., Gillison M.L., Tong B.C., Sidransky D., Califano J.A. RealTime quantitative PCR demonstrates low prevalence of human papillomavirus type 16 in
premalignant and malignant lesions of the oral cavity. Clin. Cancer Res. 2002;8(5):12031209.
[PubMed]
32. Hayashi F., Okada M., Zhong X., Miura K. PCR deteccion of Capnocytophaga species in
dental plaque samples from children aged 2 to 12 years. Microbiol. Immunol. 2001;45(1):1722.
[PubMed]
33. Heid C.A., Stevens J., Livak K.J., Williams P.M. Genome Res. 1996;6(10):986994.
[PubMed]
34. Helenius M.A., Saramaki O.R., Linja M.J., Tammela T.L., Visakorp T. Amplification of
urokinase gene in prostate cancer. Cancer Res. 2001;61(14):53405344. [PubMed]
35. Heuer H., Krsek M., Baker P., Smalla K., Wellington E.M.H. Analysis of actinomycete
communities by specific amplification of genes encoding 16S rRNA and gel-electrophoretic
separation in denaturing gradients. Appl. Environ. Microbiol. 1997;63(8):32333241. [PMC free
article] [PubMed]
36. Helweg-Larsen J., Jensen J.S., Benfield T., Svendesen U.G., Lundgren J.D., Lundgren B.
Diagnostic use of PCR for detection of Pneumocystis carinii in oral wash samples. J. Clin.
Microbiol. 1998;36(7):20682062. [PMC free article] [PubMed]
37. Helweg-Larsen J., Jensen J.S., Dohn B., Benfield T., Svendesen U.G., Lundgren B. Detection
of Pneumocystis DNA in samples from patients suspected of bacterial pneumonia- a case control
study. BMC Infect. Dis. 2002;25(2):28. [PMC free article] [PubMed]
38. Hirasawa M., Takada K. A new selective medium for Streptococcus mutans and the
distribution of S. mutans and S. sobrinus and their serotypes in dental plaque. Caries Res.
2003;37(2):212217. [PubMed]
39. Hodinka R.L. The clinical utility of viral quantitation using molecular methods. Clin. Diagn.
Virol. 1998;10(1):2547. [PubMed]
40. Huijsdens X.W., Linskens R.K., Mak M., Meuwissen S.G., Vandenbroucke-Grauls C.M.,
Savelkoul P.H. Quantification of bacteria adherent to gastrointestinal mucosa by real-time PCR.
J. Clin. Microbiol. 20002;40(12):44234427. [PMC free article] [PubMed]

41. Igarashi T., Ichikawa K., Yamamoto A., Goto N. Identification of mutans streptococcal
species by the PCR products of the dex genes. J. Microbiol. Methods. 2001;46(2):99105.
[PubMed]
42. Igarashi T., Yano Y., Yamamoto A., Sasa R., Goto N. Identification of Streptococcus
salivarius by PCR and DNA probe. Lett Appl. Microbiol. 2001;32(6):394397. [PubMed]
43. Igarashi T., Yamamoto A., Goto N. Rapid identification of mutans streptococcal species.
Microbiol. Immunol. 1996;40(11):867871. [PubMed]
44. Jordens J.Z., Lanham S., Pickett M.A., Amarasekara S., Aberywickrema I., Watt P.J.
Amplification with molecular beacon primers and reverse line blotting for the detection and
typing of human papillomaviruses. J. Virol. Methods. 2000;89(1)(2):2937. [PubMed]
45. Kariyazono H., Ohno T., Ihara K., Igareshi H., Joh-O K., Ishikawa S., et al. Rapid detection
of the 22q 11.2 deletion with quantitative real-time PCR. Mol. Cell. Probes. 2001;15(2):7173.
[PubMed]
46. Kearns A.M., Turner A.J.L., Taylor C.E., George P.W., Freeman R., Gennery A.R.
LightCycler-based quantitative PCR for rapid detection of human herpes virus 6 DNA in clinical
material. J. Clin. Microbiol. 2001;39(8):30203021. [PMC free article] [PubMed]
47. Kim Y., Flynn T.R., Donoff R.B., Wong D.T.W., Tood R. The gene: the polymerase chain
reaction and its clinical application. J. Oral Maxillofac. Surg. 2002;60(7):808815. [PubMed]
48. Kimura S., Ooshima T., Takiguchi M., Sasaki Y., Amano A., Morisaki I., et al.
Periodontopathic bacterial infection in childhood. J. Periodontol. 2002;73(1):2026. [PubMed]
49. Kostrikis L.G., Tyagi S.M., Mhlanga M., Ho D.D., Kramer F.R. Molecular Beacons: Spectral
Genotyping of Human Alleles. Science. 1998;279(5354):12281229. [PubMed]
50. Kubista M., Andrade J.M., Bengtsson M., Forootan A., Jonk J., Lind K., Sindelka R.,
Sjoback R., Sjogreen B., Strombom L., Stahlberg A., Zoric N. The real-time polymerase chain
reaction. Mol. Aspects Med. 2006;27(2-3):95115. [PubMed]
51. Kutyavin I.V., Afonina I.A., Mills A., Gorn V.V., Luckhtanov A., Belousov E.S., Singer M.J.,
Walburger D.K., Lokhov S.G., Gall A.A., Dempcy R., Reed M.W., Meyer R.B., Hedgpeth J. 3minor groove binder - DNA probes increase sequence specificity at PCR extension temperatures.
Nucleic Acids Res. 2000;28(2):65561. [PMC free article] [PubMed]
52. lee b.k., diebel e., neukam f.w., wiltfang j., ries j. Diagnostic and prognostic relevance of
expression of human telomerase subunits in oral cancer. Int. J. Oncol. 2001;19(5):10631068.
[PubMed]
53. Lehmann U., Glockner S., Kleeberger W., Feist H., Von Wasielewski R., Kreipe H. Detection
of gene amplification in archival breast cancer specimens by laser-assisted microdissection and
quantitative real-time polymerase chain reaction. Am. J. Pathol. 2000;156(6):18551864. [PMC
free article] [PubMed]
54. Lyons S.R., Griffen A.L., Leys E.J. Quantitative real-time PCR for Porphyromonas
gingivalis and total bacteria. J. Clin. Microbiol. 2000;38(6):23622365. [PMC free article]
[PubMed]
55. Lo Y.M., Chan A.T., Chan L.Y., Leung S.F., Lam C.W., Huang D.P., Johnson P.J. Molecular
prognostication of nasopharyngeal carcinoma by quantitative analysis of circulating Epstein-Barr
virus DNA. Cancer Res. 2000;60(24):68786881. [PubMed]
56. Mackay I.M., Arden K.E., Nitsche A. Real-time PCR in virology. Nucl. Ac. Res.
2002;30(6):12921305. [PMC free article] [PubMed]
57. Mauchline T.H., Kerry B.R., Hirsch P.R. Quantification in soil and the rhizosphere of the
nematophagous fungus Verticillium chamydosporium by competitive PCR and comparison with

selective plating. Appl. Environ. Microbiol. 2002;68(4):18461853. [PMC free article]

[PubMed]
58. Matsuki T., Watanabe K., Fujimoto J., Kado Y., Takada T., Matsumoto K., Tanaka R.
Quantitative PCR with 16S rRNAgene-targeted species-specific primers for analysis of human
intestinal bifidobacteria. Appl. Environ. Microbiol. 2004;70(1):167173. [PMC free article]
[PubMed]
59. Matto J., Saarela M., Alauusua S., Oja V., Jousimies-Somer H., Asikainen S. Detection of
Porphyromonas gingivalis from saliva by PCR by using a simple sample-processing method. J.
Clin. Microbiol. 1998;36(1):157160. [PMC free article] [PubMed]
60. Maccartney H.A., Foster S.J., Fraaue B.A., Ward E. Molecular diagnostics for fungal plant
pathogens. Pest. Manag. Sci. 2003;59(2):129142. [PubMed]
61. Mitas M., Mikhitarian K., Walters C., Baron P.L., Elliot B.M., Brothers T.E., Robinson J.G.,
Metcalf J.S., Palesch Y.Y., Zhang Z., Gillanders W.E., Cole D.J. Quantitative real-time RTPCR
detection of breast cancer micrometastasis using a multigene marker panel. Int. J. Cancer.
2001;93(2):162171. [PubMed]
62. Morillo J.M., Lau L., Sanz M., Herrera D., Silva A. Quantitative real time PCR based on
single copy gene sequence for detection of Actinobacillus actinomycetemcomitans and
Porphyromonas gingivalis. J. Periodont. Res. 2003;38(5):518524. [PubMed]
63. Morita E., Narikiyo M., Yano A., Nishimura E., Igaki H., Sasaki H., Terada M., Hanada N.,
Kawabe R. Different frequencies of Streptococcus anginosus infection in oral cancer and
esophageal cancer. Cancer Sci. 2003;94(6):492496. [PubMed]
64. Morrison T.B., Weis J.J., Wittwer C.T. Quantification of low-copy transcripts by continuous
SYBR Green I monitoring during amplification. BioTechniques. 1998;24(6):954958. 960962.
[PubMed]
65. Mullally B.H., Dace B., Shelburne C.E., Wolff L.F., Coulter W.A. Prevalence of periodontal
pathogens in localized and generalized forms of early-onset periodontitis. J. Periodont. Res.
2000;35(4):232241. [PubMed]
66. Mullis K.B. Target amplification for DNA analysis by the polymerase chain reaction. Ann.
Biol. Clin. 1990;48(8):579582. [PubMed]
67. Murdoch-Kinch C.A. Oral medicine: advances in diagnostic procedures. J. Calif. Dent.
Assoc. 1999;27(10):773784. [PubMed]
68. Nakanishi H., Kodera Y., Yamamura Y., Ito S., Kato T., Ezaki T., Tatematsu M. Rapid
quantitative detection of carcinoembryonic antigen-expressing free tumor cells in the peritoneal
cavity of gastric-cancer patients with real-time RT-PCR on the lightcycler. Int. J. Cancer.
2000;89(5):411417. [PubMed]
69. Nakano K., Nomura R., Nakagawa I., Hamada S., Ooshima T. Demonstration of
Streptococcus mutans with a cell wall polysaccharide specific to a new serotype K, in the human
oral cavity. J. Clin. Microbiol. 2004;42(1):198202. [PMC free article] [PubMed]
70. Nazarenko I.A., Bhatnagar S.K., Hohman R.J. A closed tube format for amplification and
detection of DNA based on energy transfer. Nucleic Acids Res. 1997;25(12):25162521. [PMC
free article] [PubMed]
71. Niesters H.G.M. Quantitation of viral load using real- time amplification techniques.
Methods. 2001;25(4):419429. [PubMed]
72. Niesters H.G.M. Molecular and diagnostic clinical virology in real time. Clin. Microbiol.
Infect. 2004;10(1):511. [PubMed]

73. Novais C.M., Pires-Alves M., Silva F.F. PCR em tempo real. Rev. Biotecnol. Cienc. Des.
2004:33.
74. Nygren J., Svanvik N., Kubista M. The interaction between the fluorescent dye thiazole
orange and DNA. Biopolymers. 1998;46(1):3951. [PubMed]
75. Okada M., Hayashi F., Nagasaka N. Detection of Actinobacillus actinomicetemcomitans and
Porphyromonas gingivalis in dental plaque samples from children 2 to 12 years of age. J. Clin.
Periodontol. 2000;27(10):763768. [PubMed]
76. Okada M., Soda Y., Hayashi F., Doi T., Suzuki J., Miura K., et al. PCR detection of
Streptococcus mutans and S. sobrinus in dental plaque samples from Japanese pr-school
children. J. Med. Microbiol. 2002;51(5):443447. [PubMed]
77. Parra B., Slots J. Detection of human viruses in periodontal pockets using polymerase chain
reaction. Oral Microbiol. Immunol. 1996;11(5):289293. [PubMed]
78. Peter M., Gilbert E., Delattre O. A multiplex real-time pcr assay for the detection of gene
fusions observed in solid tumors. Lab. Invest. 2001;81(6):905912. [PubMed]
79. Quint W.G.V., Heijtink R.A., Schirm J., Gerlich W.H., Niesters H.G.M. Reliability of
methods for hepatitis B virus DNA detection. J. Clin. Microbiol. 1995;33(1):225228. [PMC free
article] [PubMed]
80. Ranjard L., Poly F., Lata J.C., Mougel C., Thioulouse J., Nazaret S. Characterization of
bacterial and fungal soil communities by automated ribosomal intergenic spacer analysis
fingerprints: Biological e Methodological variability. Appl. Environ. Microbiol.
2001;67(10):44794487. [PMC free article] [PubMed]
81. Reis P.P., Rogatto S.R., Kowalski L.P., Nishimoto I.N., Montovani S.C., Corpus G., et al.
Quantitative real-time PCR identifies a critical region of deletion on 22q13 related to prognosis
in oral cancer. Oncogene. 2002;21(42):64806487. [PubMed]
82. Richard B., Groisillier A., Badet C., Dorignac G., Lonvaud-Funel A. Identification of
salivary Lactobacillus rhamnosus species by DNA profiling and a specific probe. Res.
Microbiol. 2001;152(2):157165. [PubMed]
83. Riggio M.P., Lennon A. Rapid identification of Actinobacillus actinomycetemcomitans,
Haemophilus aphrophilus, and Haemophilus paraphrophilus by restriction enzyme analysis of
PCR-amplified 16S rRNA genes. J. Clin. Microbiol. 1997;35(6):16301632. [PMC free article]
[PubMed]
84. Ririe K.M., Rasmussen R.P., Wittwer C.T. Product differentiation by analysis of DNA
melting curves during the polymerase chain reaction. Anal. Biochem. 1997;245(2):154160.
[PubMed]
85. Ras I.N. Polymerase Chain Reaction-PCR. In: LOPES H.P., SIQUEIRA Jr., editors.
Endodontia:Biologia e Tcnica. 2 ed. Rio de Janeiro: Guanabara Koogan; 2004. Box 10-1.
86. Rupf S., Kneist S., Merte K., Eschrich K. Quantitative determination of Streptococcus
mutans by using competitive polymerase chain reaction. Eur. J. Oral Sci. 1999;107(2):7581.
[PubMed]
87. Saarela M., Hannula J., Matto J., Asikainen S., Alaluusua S. Typing of mutans streptococci
by arbitrarily primed polymerase chain reaction. Arch. Oral Biol. 1996;41(8-9):821826.
[PubMed]
88. Sakamoto M., Takeuchi Y., Umeda M., Ishikawa I., Benno Y. Rapid deteccion and
quantification of live periodontopathic bacteria by real time PCR. Microbiol. Immunol.
2001;45(1):3944. [PubMed]

89. Sakamoto M., Takeuchi Y., Umeda M., Ishikawa I., Benno Y., Nakase T. Detection of
Treponema socranskii associated with human periodontitis by PCR. Microbiol. Immunol.
1999;43(5):485490. [PubMed]
90. Santos C.F., Sakai V.T., Machado M.A.A.M., Schippers D.N., Greene A.S. Reverse
transcription and polymerase chain reaction: principles and applications in dentistry. J. Appl.
Oral Sci. 2004;12(1):111. [PubMed]
91. Saygun I., Sahin S., Ozdemir A., Kurtis B., Yapar M., Kubar A., et al. Detection of human
viruses in patients with chronic periodontitis and the relationship between viruses and clinical
parameters. J. Periodontol. 2002;73(12):14371443. [PubMed]
92. Schuurman R., Descamps D., Weverling G.J., Kaye S., Tijnagel J., Williams I., Van Leeuwen
R., Tedder R., Boucher C.A., Brun-Vezinet F., Loveday C. Multicenter comparison of three
commercial methods for quantification of human immunodeficiency virus type 1 RNA in
plasma. J. Clin. Microbiol. 1996;34(12):30163022. [PMC free article] [PubMed]
93. Shelburne C.E., Gleason R.M., Germaine G.R., Wolff L.F., Mullally B.H., Coulter W.A., et
al. Quantitative reverse transcription polymerase chain reaction analysis of Porphyromonas
gingivalis gene expression in vivo. J. Microbiol. Meth. 2002;49(2):147156. [PubMed]
94. Silva L.A.F., Passos N.S. Coleta de amostras biolgicas em locais de crimes para estudo de
DNA. Macei: UFAL; 2002. DNA Forense.
95. Siqueira Jr. J.F., Ras I.N. A 16 rDNA-based nested PCR protocol to detect Campylobacte
gracilis in oral infecctions. Pesqui. Odontol. Bras. 2003;17(2):142146. [PubMed]
96. Siqueira Jr. J.F., Ras I.N. PCR methodology as a valuable tool for identification of
endodontic pathogens. J. Dent. 2003;31(5):333339. [PubMed]
97. Slots J. Rapid identification of important periodontal microorganisms by cultivation. Oral
Microbiol. Immunol. 1986;1(1):4857. [PubMed]
98. Song Y. PCR-based diagnostics for anaerobic infections. Anaerobe. 2005;11(1-2):7991.
[PubMed]
99. Speers D.J., Ryan S., Harnett G., Chidlow G. Diagnosis of malaria aided by polymerase
chain reaction in two cases with low-level parasitaemia. Inter. Med. J. 2003;33(12):613615.
[PubMed]
100. Speers D.J. Clinical applications of molecular biology for infectious diseases. Clin.
Biochem. Rev. 2006;27(1):3950. [PMC free article] [PubMed]
101. Spolidorio D.M.P., Spolidorio L.C. Tcnicas bsicas de biologia molecular. In: ESTRELA
C., editor. Metodologia cientfica-Cincia-Ensino-Pesquisa. So Paulo: Artes Mdicas; 2005.
102. Sterflinger K., Krumbien W.E., Schwiertz A. A protocol for PCR in situ hybridization of
hypomycetes. Int. Microbiol. 1998;1(3):213220. [PubMed]
103. Takeuchi Y., Makoto U., Sakamoto M., Benno Y., Huang Y., Ishikawa I. Treponema
socranskii, Treponema denticola, and Porphyromonas gingivalis are associated with severity of
periodontal tissue destruction. J. Periodontol. 2001;72(10):13541363. [PubMed]
104. Tran S.D., Rudney J.D. Improved multiplex PCR using conserved and species-specific 16S
rRNA gene primer for simultaneous detection. J. Clin. Microbiol. 1999;37(11):35043508.
[PMC free article] [PubMed]
105. Truong T.L., Menard C., Mouton C., Trahan L. Identification of mutans and other oral
Streptococci by random amplified polymorphic DNA analysis. J. Med. Microbiol.
2000;49(1):6371. [PubMed]
106. Tyagi S., Kramer F.R. Molecular beacons: probes that fluoresce upon hybridization. Nat.
Biotechnol. 1996;14(3):303308. [PubMed]

107. Tyagi S., Bratu D.P., Kramer F.R. Multicolor molecular beacons for allele discrimination.
Nat. Biotechnol. 1998;16(1):4953. [PubMed]
108. Whiley R.A., Beighton D. Current classification of the oral streptococci. Oral Microbiol.
Immunol. 1998;13(4):195216. [PubMed]
109. Whitcombe D., Brownie J., Gillard H.L., Mckechnie D., Theaker J., Newton C.R., Little S.
A homogeneous fluorescence assay for PCR amplicons: its application to real-time single-tube
genotyping. Clin. Chem. 1998;44(5):918923. [PubMed]
110. Whitcombe D., Theaker J., Guy S.P., Brown T., Little S. Detection of PCR products using
self probing amplicons and fluorescence. Nat. Biotechnol. 1999;17(8):804807. [PubMed]
111. Williamson E.M.C., Leeming J.P., Palmer H.M., Steward C.G., Warnock D., Marks D.I.,
Millar M.R. Diagnosis of invasive aspergillosis in bone marrow transplant recipients by
polymerase chain reaction. Br. J. Haematol. 2000;108(1):132139. [PubMed]
112. Winton L.M., Stone J.K., Watrud L.S., Hansen E.M. Simultaneous one-tube quantification
of host and pathogen DNA with real-time polymerase chain reaction. Phytopathology.
2002;92:102116. [PubMed]
113. Wittwer C.T., Herrmann M.G., Moss A.A., Rasmussen R.P. BioTechniques.
1997;22(1):130131. 134138. [PubMed]
114. Wolk D., Mitchell P.S., Patel R. Principles of molecular microbiology testing methods. In:
COCKERILL F., editor. Infectious Diseases Clinics of North America. ed. Philadelphia: WB
Saunders; 2001. [PubMed]
115. Wu P., Brand L. Resonance energy transfer: methods and applications. Anal.Biochem.
1994;218(1):113. [PubMed]
116. Yang S., Rothman R.E. PCR-based diagnostics for infectious diseases: uses, limitations, and
future applications in acute-care settings. Lancet Infect. Dis. 2004;4(6):337348. [PubMed]
117. Yoo S.Y., Kim P.S., Hwang H.K., Lim S.H., Kim K.W., Choe S.J., Min B.M., Kook J.K.
Identification of non-mutans Streptococci organisms in dental plaques recovering on
mitissalivarius bacitracin agar medium. J. Microbiol. 2005;43(2):204208. [PubMed]
118. YOSHIDA A., SUZUKI N., NAKANO Y., KAWADA M., OHO T., KOGA T. Development
of a 5 nuclease-based real-time PCR assay for quantitative detection of cariogenic dental
pathogens Streptococcus mutans and Streptococcus sobrinus. J. Clin. Microbiol.
2003;41(9):44384441. [PMC free article] [PubMed]
119. Zariffard M.R., Saifuddin M., Sha B.E., Spear G.T. Detection of bacterial vaginosis-related
organisms by real-time PCR for Lactobacilli, Gardnerella vaginalis and Mycoplasma hominis.
FEMS Immunol. Med. Microbiol. 2002;34(4):277281. [PubMed]
120. Zerr D.M., Huang M.L., Corey L., Erickson M., Parker H.L., Frenkel L.M. Sensitive
method for detection of human herpesvirus 6 and 7 in saliva collected in field studies. J. Clin.
Microbiol. 2000;38(5):19811983. [PMC free article] [PubMed]
Articles from Brazilian Journal of Microbiology are provided here courtesy of Elsevier

Formats:

Article
|

PubReader
|

ePub (beta)
|

PDF (180K)
|

Citation

Share

Facebook

Twitter

Google+

Save items
View more options

Similar articles in PubMed

Rapid detection, characterization, and enumeration of foodborne pathogens.[APMIS

Suppl. 2011]

Principles and applications of polymerase chain reaction: basic science for the practicing
physician.[Ann Allergy Asthma Immunol. 2008]

Fungal molecular diagnostics: a mini review.[J Appl Genet. 2004]

Rapid detection and limitations of molecular techniques.[Annu Rev Food Sci Technol.
2011]

Real-Time PCR: Revolutionizing Detection and Expression Analysis of Genes.[Curr

Genomics. 2007]

See reviews...See all...

Cited by other articles in PMC

Polymerase chain reaction: A molecular diagnostic tool in periodontology[Journal of

Indian Society of P...]

See all...

Links

PubMed

Taxonomy

Recent Activity
ClearTurn Off

Principles and applications of polymerase chain reaction in medical diagnostic f...

Principles and applications of polymerase chain reaction in medical diagnostic fields: a
review
Brazilian Journal of Microbiology. Jan-Mar 2009; 40(1)1

See more...

Target amplification for DNA analysis by the polymerase chain reaction.[Ann Biol Clin
(Paris). 1990]

Review Fungal molecular diagnostics: a mini review.[J Appl Genet. 2004]

See more ...

Quantitative real-time PCR based on single copy gene sequence for detection of
Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis.[J Periodontal
Res. 2003]

Review The real-time polymerase chain reaction.[Mol Aspects Med. 2006]

Quantification of low-copy transcripts by continuous SYBR Green I monitoring during

amplification.[Biotechniques. 1998]

Continuous fluorescence monitoring of rapid cycle DNA amplification.[Biotechniques.

1997]

The interactions between the fluorescent dye thiazole orange and DNA.[Biopolymers.
1998]

Product differentiation by analysis of DNA melting curves during the polymerase chain
reaction.[Anal Biochem. 1997]

3'-minor groove binder-DNA probes increase sequence specificity at PCR extension

temperatures.[Nucleic Acids Res. 2000]

Detection of nucleic acid hybridization by nonradiative fluorescence resonance energy

transfer.[Proc Natl Acad Sci U S A. 1988]

Review Resonance energy transfer: methods and applications.[Anal Biochem. 1994]

Homogeneous amplification and variant detection by fluorescent hybridization probes.

[Clin Chem. 2000]

A novel method for real time quantitative RT-PCR.[Genome Res. 1996]

Real time quantitative PCR.[Genome Res. 1996]

Molecular beacons: probes that fluoresce upon hybridization.[Nat Biotechnol. 1996]

Multicolor molecular beacons for allele discrimination.[Nat Biotechnol. 1998]

Spectral genotyping of human alleles.[Science. 1998]

Review Current classification of the oral streptococci.[Oral Microbiol Immunol. 1998]

A closed tube format for amplification and detection of DNA based on energy transfer.
[Nucleic Acids Res. 1997]

A homogeneous fluorescence assay for PCR amplicons: its application to real-time,

single-tube genotyping.[Clin Chem. 1998]

A closed tube format for amplification and detection of DNA based on energy transfer.
[Nucleic Acids Res. 1997]

Detection of PCR products using self-probing amplicons and fluorescence.[Nat

Biotechnol. 1999]

Review Real-time PCR in virology.[Nucleic Acids Res. 2002]

Review Quantitation of viral load using real-time amplification techniques.[Methods.

2001]

Review The clinical utility of viral quantitation using molecular methods.[Clin Diagn
Virol. 1998]

Review Molecular and diagnostic clinical virology in real time.[Clin Microbiol Infect.
2004]

Multicenter comparison of three commercial methods for quantification of human

immunodeficiency virus type 1 RNA in plasma.[J Clin Microbiol. 1996]

Reliability of methods for hepatitis B virus DNA detection.[J Clin Microbiol. 1995]

See more ...

LightCycler-based quantitative PCR for rapid detection of human herpesvirus 6 DNA in

clinical material.[J Clin Microbiol. 2001]

Sensitive method for detection of human herpesviruses 6 and 7 in saliva collected in field
studies.[J Clin Microbiol. 2000]

Review Real-time PCR in virology.[Nucleic Acids Res. 2002]

See more ...

Review Real-time PCR in virology.[Nucleic Acids Res. 2002]

Quantitation of varicella-zoster virus DNA in patients with Ramsay Hunt syndrome and
zoster sine herpete.[J Clin Microbiol. 2001]

LightCycler-based quantitative PCR for rapid detection of human herpesvirus 6 DNA in

clinical material.[J Clin Microbiol. 2001]

See more ...

Clinical applications of molecular biology for infectious diseases.[Clin Biochem Rev.

2006]

Review Molecular diagnostics for fungal plant pathogens.[Pest Manag Sci. 2003]

Review Fungal molecular diagnostics: a mini review.[J Appl Genet. 2004]

A protocol for PCR in situ hybridization of hyphomycetes.[Int Microbiol. 1998]

Review Molecular community analysis of microbial diversity.[Curr Opin Biotechnol.

2002]

See more ...

Diagnostic use of PCR for detection of Pneumocystis carinii in oral wash samples.[J Clin
Microbiol. 1998]

Detection of Pneumocystis DNA in samples from patients suspected of bacterial

pneumonia--a case-control study.[BMC Infect Dis. 2002]

Diagnosis of invasive aspergillosis in bone marrow transplant recipients by polymerase

chain reaction.[Br J Haematol. 2000]

The application of real-time PCR to the identification, detection and quantification of

Pyrenophora species in barley seed.[Mol Plant Pathol. 2001]

See more ...

Diagnosis of malaria aided by polymerase chain reaction in two cases with low-level
parasitaemia.[Intern Med J. 2003]

Review Principles of molecular microbiology testing methods.[Infect Dis Clin North Am.
2001]

Detection of Bacillus anthracis DNA by LightCycler PCR.[J Clin Microbiol. 2002]

Detection of smallpox virus DNA by LightCycler PCR.[J Clin Microbiol. 2002]

Application of rapid-cycle real-time polymerase chain reaction for diagnostic testing in

the clinical microbiology laboratory.[Arch Pathol Lab Med. 2003]

PCR-based diagnostics for anaerobic infections.[Anaerobe. 2005]

Review PCR-based diagnostics for infectious diseases: uses, limitations, and future
applications in acute-care settings.[Lancet Infect Dis. 2004]

A survey of antimicrobial susceptibility testing in the United Kingdom.[J Antimicrob

Chemother. 1996]

See more ...

Rapid identification of important periodontal microorganisms by cultivation.[Oral

Microbiol Immunol. 1986]

Review PCR-based diagnostics for infectious diseases: uses, limitations, and future
applications in acute-care settings.[Lancet Infect Dis. 2004]

Detection of bacterial vaginosis-related organisms by real-time PCR for Lactobacilli,

Gardnerella vaginalis and Mycoplasma hominis.[FEMS Immunol Med Microbiol. 2002]

See more ...

Review PCR-based diagnostics for infectious diseases: uses, limitations, and future
applications in acute-care settings.[Lancet Infect Dis. 2004]

Clinical applications of molecular biology for infectious diseases.[Clin Biochem Rev.

2006]

Review The gene: the polymerase chain reaction and its clinical application.[J Oral
Maxillofac Surg. 2002]

Identification of salivary Lactobacillus rhamnosus species by DNA profiling and a

specific probe.[Res Microbiol. 2001]

Rapid detection and quantification of five periodontopathic bacteria by real-time PCR.

[Microbiol Immunol. 2001]

Treponema socranskii, Treponema denticola, and Porphyromonas gingivalis are

associated with severity of periodontal tissue destruction.[J Periodontol. 2001]

PCR detection of Capnocytophaga species in dental plaque samples from children aged 2
to 12 years.[Microbiol Immunol. 2001]

See more ...

Reverse transcription and polymerase chain reaction: principles and applications in

dentistry.[J Appl Oral Sci. 2004]

Polymerase chain reaction detection of 8 putative periodontal pathogens in subgingival

plaque of gingivitis and advanced periodontitis lesions.[Oral Microbiol Immunol. 1996]

See more ...

Enumeration of Porphyromonas gingivalis, Prevotella intermedia and Actinobacillus

actinomycetemcomitans in subgingival plaque samples by a quantitative-competitive
PCR method.[J Med Microbiol. 2000]

Rapid identification of Actinobacillus actinomycetemcomitans, Haemophilus

aphrophilus, and Haemophilus paraphrophilus by restriction enzyme analysis of PCRamplified 16S rRNA genes.[J Clin Microbiol. 1997]

Detection of human viruses in periodontal pockets using polymerase chain reaction.[Oral

Microbiol Immunol. 1996]

Detection of human viruses in patients with chronic periodontitis and the relationship
between viruses and clinical parameters.[J Periodontol. 2002]

Quantitative reverse transcription polymerase chain reaction analysis of Porphyromonas

gingivalis gene expression in vivo.[J Microbiol Methods. 2002]

Detection of Porphyromonas gingivalis from saliva by PCR by using a simple sampleprocessing method.[J Clin Microbiol. 1998]

Detection of Treponema socranskii associated with human periodontitis by PCR.

[Microbiol Immunol. 1999]

Prevalence of periodontal pathogens in localized and generalized forms of early-onset

periodontitis.[J Periodontal Res. 2000]

Improved multiplex PCR using conserved and species-specific 16S rRNA gene primers
for simultaneous detection of Actinobacillus actinomycetemcomitans, Bacteroides
forsythus, and Porphyromonas gingivalis.[J Clin Microbiol. 1999]

Detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in

dental plaque samples from children 2 to 12 years of age.[J Clin Periodontol. 2000]

Quantitative real-time PCR for Porphyromonas gingivalis and total bacteria.[J Clin
Microbiol. 2000]

Rapid detection and quantification of five periodontopathic bacteria by real-time PCR.

[Microbiol Immunol. 2001]

Review The gene: the polymerase chain reaction and its clinical application.[J Oral
Maxillofac Surg. 2002]

See more ...

Demonstration of Streptococcus mutans with a cell wall polysaccharide specific to a new

serotype, k, in the human oral cavity.[J Clin Microbiol. 2004]

Review Current classification of the oral streptococci.[Oral Microbiol Immunol. 1998]

A new selective medium for Streptococcus mutans and the distribution of S. mutans and
S. sobrinus and their serotypes in dental plaque.[Caries Res. 2003]

Identification of non-mutans streptococci organisms in dental plaques recovering on

mitis-salivarius bacitracin agar medium.[J Microbiol. 2005]

See more ...

Identification of salivary Lactobacillus rhamnosus species by DNA profiling and a

specific probe.[Res Microbiol. 2001]

Rapid identification of mutans streptococcal species.[Microbiol Immunol. 1996]

Typing of mutans streptococci by arbitrarily primed polymerase chain reaction.[Arch

Oral Biol. 1996]

Quantitative determination of Streptococcus mutans by using competitive polymerase

chain reaction.[Eur J Oral Sci. 1999]

PCR methodology as a valuable tool for identification of endodontic pathogens.[J Dent.

2003]

Review The gene: the polymerase chain reaction and its clinical application.[J Oral
Maxillofac Surg. 2002]

Black-pigmented anaerobic rods in closed periapical lesions.[Int Endod J. 1999]

Rapid identification of important periodontal microorganisms by cultivation.[Oral

Microbiol Immunol. 1986]

Occurrence of Candida albicans in infections of endodontic origin.[J Endod. 2000]

A 16S rDNA-based nested PCR protocol to detect Campylobacter gracilis in oral

infections.[Pesqui Odontol Bras. 2003]

Rapid detection of the 22q11.2 deletion with quantitative real-time PCR.[Mol Cell
Probes. 2001]

See more ...

Different frequencies of Streptococcus anginosus infection in oral cancer and esophageal

cancer.[Cancer Sci. 2003]

Genetic progression model for head and neck cancer: implications for field cancerization.
[Cancer Res. 1996]

Evidence for a causal association between human papillomavirus and a subset of head
and neck cancers.[J Natl Cancer Inst. 2000]

Real-time quantitative PCR demonstrates low prevalence of human papillomavirus type

16 in premalignant and malignant lesions of the oral cavity.[Clin Cancer Res. 2002]

Novel approach to quantitative polymerase chain reaction using real-time detection:

application to the detection of gene amplification in breast cancer.[Int J Cancer. 1998]

Detection of gene amplification in archival breast cancer specimens by laser-assisted

microdissection and quantitative real-time polymerase chain reaction.[Am J Pathol. 2000]

See more ...

You are here: NCBI > Literature > PubMed Central (PMC)
Write to the Help Desk
External link. Please review our privacy policy.
NLM
NIH
DHHS
USA.gov
Copyright | Disclaimer | Privacy | Browsers | Accessibility | Contact
National Center for Biotechnology Information, U.S. National Library of Medicine 8600
Rockville Pike, Bethesda MD, 20894 USA