Infection, Genetics and Evolution 2 (2002) 153–158

Discussion
Molecular epidemiology of Mycobacterium tuberculosis and its
relevance to the surveillance and control of TB: an e-debate
Loubna Tazi a , Barry Kreiswirth b , Christian Carrière c , Michel Tibayrenc a,∗
a Genetics of Infectious Diseases, Unité mixte de Recherche Centre National de la Recherch Scientifique/Institut de Recherche
pour le Développement no. 9926, IRD, BP 64501, 34394 Montpellier Cedex 5, France
b Public Health Research Institute Tuberculosis Center, International Center for Public Health,

225 Warren Street, Newark, NJ 07103, USA

c Laboratoire de Bactériologie, Hôpital Arnaud de Villeneuve, 371 Avenue du Doyen Giraud,

34295 Montpellier Cedex 5, France

Accepted 19 September 2002

1. First question: Give in a few lines your definition of • and the last but not the least, to develop strategies for the
‘Molecular Epidemiology’ and your opinion about what treatment and the prevention of the disease.
it is good for in general (not only for TB)
1.2. Response from BK
1.1. Response from LT
As correctly described by Loubna, molecular epidemi-
In the last 20 years, Molecular Epidemiology has be- ology has opened many pathways of investigation and in
come a major field of research, which corresponds to the regard to tuberculosis, a number of central dogmas have
integration of molecular approaches into the conventional been challenged. This includes the finding that exogenous
epidemiologic studies. This scientific domain implies sev- infections occur in both HIV+ and HIV− tuberculosis pa-
eral disciplines, comprising medicine, molecular biology, tients, and that the percentage of recent transmission in a
epidemiology and biostatistics. given population is higher than previously expected. The
Molecular epidemiology can be defined as a science that molecular tools have also provided precise markers to dis-
permits to understand the transmission, the pathogenesis and tinguish between primary and acquired drug resistance in
the etiology of a disease in a human populations. a given population. In regard to virulence and pathogenesis
Based on this definition, the use of molecular epidemiol- studies, molecular epidemiology proves to be directive in
ogy to study the infectious diseases has mainly permitted to identifying clones associated with a given phenotype—from
or will permit to: drug resistance to toxin production.
• estimate the association of risk factors with the transmis- It should be stressed that the information derived from
sion of the disease; molecular epidemiological investigations is always depen-
• detect and confirm outbreaks in institutional settings; dent on the sampling and that a non-bias, representative col-
• screen strains of public health importance; lection is critical to each study.
• distinguish between reactivation and recent infection (case
of TB); 1.3. Response from CC
• track global spread of pathogens;
• understand the virulence and resistance mechanisms of The molecular epidemiology is the application of molec-
different strains; ular biology techniques to the study of the distribution and
• improve the knowledge on transmission dynamics and determinants of an infection and communicable diseases.
dissemination pathways of infectious diseases; For the clinical microbiologist the notion of epidemiologic
problem is most often related to a nosocomial infection
which is usually caused by bacteria. To verify the existence
∗ Corresponding author. Tel.: +33-4-6741-6197; fax: +33-4-6741-6299. of a nosocomial infection it is necessary to establish that all
E-mail address: michel.tibayrenc@mpl.ird.fr (M. Tibayrenc). cases are caused by the same bacteria. That is the role of the

1567-1348/02/$ – see front matter © 2002 Elsevier Science B.V. All rights reserved.
PII: S 1 5 6 7 - 1 3 4 8 ( 0 2 ) 0 0 1 3 2 - 6
154 L. Tazi et al. / Infection, Genetics and Evolution 2 (2002) 153–158

microbiologist. When epidemiologic typing of the bacterial has had a major impact on the control and treatment of tu-
strains is performed, phenotypic techniques are most often berculosis.
insufficient for discriminating among the isolates. The use
of molecular techniques gives most of the time the answer
about whether strains are epidemiologically related or not. 2.2. Response from CC

The case of cross-contamination of clinical samples

1.4. Comment from MT during the sampling or culture procedure is possible and
more and more frequently reported (Bauer et al., 1997;
About Loubna’s excellent response: please remember that
Gascoyne-Binzi et al., 2001; Ramose et al., 1999). Thanks
the interest of molecular epidmeiology is not limited to
to molecular epidemiology study of the Mycobacterium
human infections, and includes pathogens of animals and
tuberculosis isolates these incidents can be identify.
plants. This makes it possible, in the domain of human
The source of cross-contamination can be the sampling
medicine, to clarify the epidemiology of human diseases that
material (contamined bronchoscope) or laboratory contami-
have an animal reservoir, such as Chagas disease (caused by
nation of cultures. In the first case, the material has not been
the parasitic protozoan Trypanosoma cruzi, that can infect
sterilized properly (error in the decontamination procedure).
any mammal) or infections by Mycobacterium bovis.
In the second case, patient’s culture is positive for M. tuber-
culosis but there is not a high clinical suspicion for disease;
the culture contamination involves positive clinical samples
2. Second question: In the field of tuberculosis, have processed concurrently with negative samples.
you heard about, or can you think about—a case where These errors can lead to unnecessary visits of falsely di-
a result from a molecular epidemiology survey has led, agnosed patients to medical consultants and unnecessary
or could lead, to a practical decision in terms of public long-term antimicrobial treatment. In terms of public health,
health? mycobacteriology laboratories should institute strict proce-
dures to prevent and identify cross-contamination episodes.
2.1. Response from BK
Careful documentation of positive specimens, together with
Over the last decade the genotyping of M. tuberculosis close liaison between clinical and laboratory staff involved
has provided evidence based data to evaluate transmission, in culturing and molecular characterization, is necessary to
distinguishing relapse from exogenous reinfection, and in identify such incidents effectively.
identifying cases of laboratory cross-contamination. The fact
that patients are subjected to incorrect medical decisions
based on a false laboratory result is a serious public health 2.3. Response from LT
problem. We and others have shown that the use of routine
monitoring of the mycobacteriology laboratory and the use Besides the case of the cross-contamination raised by
of rapid techniques, such as spoligotyping, has a positive Dr. Carrière and Dr. Kreiswirth, molecular epidemiology
impact identifying these cases. allowed also to identify and to understand nosocomial in-
In addition to laboratory cross-contamination, there are fections, exogenous reinfection, and to distinguish recent
unique settings where genotyping M. tuberculosis has had infection from reactivation of latent infection. The last case
an impact on medical care. Decisions on using pyrazinamide cited seems to me very fundamental to establish decisions
in a treatment regimen has been problematic, as phenotypic in terms of public health.
susceptibility analysis is inaccurate and provides no clini- Active tuberculosis may result from the reactivation of
cal guidance. Sequence analysis of the pncA gene and the latent infection or from recent infection. The molecular dis-
high correlation between mutations in pncA and resistance tinction between these two cases led to determine the risks
provides a laboratory basis to change treatment decisions. for TB transmission in the communities, and to strengthen
There are also rare cases where the genotype of an M. the control of this disease in function of the epidemic situ-
tuberculosis strain impacts on the treatment regimen. As ation identified.
an example, the New York City multidrug-resistant (MDR) In spite of these advances, the situation in developing
outbreak clone, strain W, which has caused disease in over countries like Morocco, my country of origin, is worrying.
500 patients, is always resistant “first-line” therapy and its Despite the recommendations of WHO (DOTS strategy), the
identification by IS6110 analysis provides strong evidence incidence of TB is still very high, and the problem of HIV
to alter therapy to second-line agents. The development of a and multidrug-resistance promotes this increasing in such
rapid PCR-based multiplex method to rapidly identify this communities. The future challenge for molecular epidemi-
primary resistant clone was developed at the CDC with the ology is to provide better understanding of the transmission
aim of controlling the spread of this serious MDR clone. dynamics of tuberculosis in these countries and to stimulate
It is clear that over the last decade, the integration of the implementation of control measures on a more global
molecular techniques in the mycobacteriology laboratory scale.
 L. Tazi et al. / Infection, Genetics and Evolution 2 (2002) 153–158 155

3. Third question: Here is a list of markers classically (ii) About RAPD: its reputation of non-reproducibility is
used in molecular epidemiology. Please expose your not justified. In experienced hands, its reproducibility is fair.
opinion about their qualities and drawbacks for I agree that it is more a research tool than a tool for routine
MTB molecular typing and their suitability for identification. However, its power in terms of population
internet-operated surveillance networks: Multilocus genetics (by allowing multilocus analysis) and phylogenetic
Enzyme Electrophoresis (MLEE); Random Primed analysis (by identifying various species- and strain-specific
Amplified Polymorphic DNA; Pulse Field Gel (“synapomorphic”) characters) is much higher than of
Electrophoresis (PFGE); Spoligotyping; IS6110 IS6110 RFLP. By its power in elucidating M. tuberculosis
Restriction Fragment Length Polymorphism (RFLP) population structure and that of many other pathogens, and
typing; Multilocus Sequence Typin (MLST) in reliably deliminating many species, its contribution to
molecular epidemiology has been indirectly considerable.
3.1. Response from BK
3.3. Comment from BK on that comment
M. tuberculosis has a monomorphic genome, similar to
the limited polymorphism observed in isolates of Bacillus Michel: The polymorphism is not only in housekeeping
anthracis or Yersinia pestis. The comparative analyses of the genes but in most genes analyzed. The Musser lab has done
available genomes and the comparative DNA sequence stud- extensive comparative sequencing on more than 50 genes
ies revealed limited synonymous mutations; a finding that led and even third position codon changes are rare. The genome
to the hypothesis that M. tuberculosis is a “young” pathogen. data on the clinical isolates supports this finding.
As a result of the limited diversity, MLEE and MLST do It is also clear that the SNP approach will be much more
not provide a high level of discrimination and are not the informative than RAPD analyses and provide a more objec-
method of choice to differentiate M. tuberculosis isolates. tive approach for both genotyping and evolutionary studies.
In considering genotyping methods to sub-speciate bacte-
ria the IS6110 system is by far the most well accepted stan- 3.4. MT
dardized method and performed on a global scale. Currently
there are more than 60,000 isolates that have been genotyped
Barry: Musser’s hypothesis has been recently challenged
with IS6110 and this data is comparable from laboratory to
by Fleischmann et al. (2002), who found an extensive poly-
laboratory. IS6110 is clearly the present gold standard with
morphism in MTB based on SNP analysis. Please note that
the understanding that it has well known limitations—strains
this perfectly fits RAPD results of our group (Loubna Tazi
with six or less copies of IS6110 should be re-evaluated us-
and Anne-Laure Bañuls).
ing an unrelated method such as spoligotyping, MIRUs or
Southern hybridization with PGRS.
PFGE and Random Primed Amplified Polymorphic DNA 3.5. Response from CC to third question
does not provide any advantage to the current use of IS6110
and spoligotyping. PFGE analysis has not been well es- The IS6110 RFLP typing is the most widely applied
tablished for M. tuberculosis, standardization and analy- molecular typing method for M. tuberculosis. To facilitate
sis between laboratories is difficult and there are questions the interlaboratory comparability of this method, all aspects
whether the procedure needs to be run in a biosafety cabinet. of the procedure have been standardized and it is considered
As with all PCR-based method, standardization of the Ran- to be the reference method for M. tuberculosis typing. How-
dom Primed Amplified Polymorphic DNA has never been ever, this technique suffers from significant drawbacks, it is
shown to be reproducible within and among laboratories laborious, requiring many technical steps and several micro-
and regardless, it will not provide the discrimination that is grams of chromosomal DNA, this implies a culture delay of
found with IS6110 and spoligotyping. a few weeks and requires viable organisms. It is an expensive
It is likely that SNP analysis will be the eventual method method and sophisticated computer software is required to
to genotype M. tuberculosis and replace IS6110 and spolig- analyze the patterns in an accurate way. On the other hand,
otyping. This will be an objective approach that will ulti- it has been possible to establish international databases of
mately database arrays of genetic alterations in numerous RFLP patterns from different geographical area. Thanks to
target loci and each strain will be defined by the collective that database it is possible to trace the source of infection
presence or absence of mutations at these sites. for multidrug-resistant strains and it permitted to know that
the Beijing genotype has a high impact on the tuberculosis
3.2. Comment from MT on BK’s response to the third epidemic in Asia and USSR republics.
question Recently, PCR-based methods have been developed. One
of them, spolygotyping is a method detecting 43 known
Barry: (i) It is important to make clear that the monomor- spacer sequences which intersperse the DRs in the ge-
phism of MTB genome concerns only housekeeping genes nomic DR region of M. tuberculosis complex strain. It is
and should be rather termed—“limited polymorphism”. easy, robust, highly reproducible and rapid. Moreover the
156 L. Tazi et al. / Infection, Genetics and Evolution 2 (2002) 153–158

results can be read as a digital code suitable for use in mits also to differentiate between members of M. tubercu-
a computer-assisted analysis and, therefore, for internet losis complex.
surveillance network. Another advantage of spolygotyping This technique is rapid and highly reproducible. More-
is that it can be used simultaneously for the detection and over, like RFLP-IS6110, a database for internet-operated
typing of the M. tuberculosis complex bacteria in one assay. surveillance networks is available. Despite all these prop-
It could be interesting to study Beijing epidemic since these erties, the spoligotyping remains less discriminative than
strains harbour a typical spolygotype pattern (reaction with RFLP-IS6110, and as RFLP-IS6110, it cannot be used for
the last nine spacers in the panel of 43). population genetics study.
However, spolygotyping has shown a discriminatory abil- Few studies have used Pulsed Field Gel Electrophoresis
ity lower than of IS6110 RFLP analysis with the exception (PFGE) for molecular typing of MTB strains. This technique
of that for M. tuberculosis isolates with low copy numbers. seems especially laborious, but PFGE analyses provide more
Another PCR-based method, the MIRU–VNTRs has been discrimination among isolates with fewer than five copies
developed. It is based on the variability in copy numbers of of IS6110 and less clustering in isolates with five or more
tandem repeats of 40–100 bp in length at 12 different inter- copies. Moreover, the application of PFGE with four inde-
genic regions of the M. tuberculosis complex genome. The pendent enzymes can be used for population genetics.
discriminatory power of the 12 MIRU–VNTR regions is Concerning Random Amplified Polymorphism DNA
much higher than that of spolygotyping and close of IS6110 (RAPD), its application on molecular epidemiology of M.
RFLP for typing of M. tuberculosis. The results can be read tuberculosis has been limited because of problems of stan-
as digital code and this opens the way to the construction of dardization of this method between laboratories. RAPD is
databases. The simple numerical format of the data gener- used in routine in our lab to type several microorganisms,
ated should allow laboratories in different parts of the world and the selection of primers seem to be fundamental, since
to compare their local isolates with those found elsewhere only the primers that give reproducible and legible patterns
by submitting their MIRU–VNTR data to a central database are used. Once the problem of reproducibility resolved,
that can be created on a web site. RAPD presents some properties: it is a rapid technique and
To my knowledge, at the present time, three databases are it requires few ADN (20 ng). Moreover, it is a multilocus
available for internet-operated surveillance network of M. marker, and it can be used for population genetics study.
tuberculosis: the RFLP patterns, the spolygotype patterns RAPD is also a generalist marker, so it permits the compar-
and the MIRU–VNTR pattern. ison of the genetic diversity among several microorganisms.
As underlined by Dr. Kreiswirth, several studies have
3.6. Response from LT to third question shown that M. tuberculosis genome presents a limited ge-
netic diversity (Sreevatsan et al., 1997). As this negligi-
The most widely used genotyping method for MTB iso- ble polymorphism concerns the structural genes, the use of
lates is the Restriction Fragment Length Polymorphism-based MLEE and MLST is not informative for the differentiation
on IS6110. This technique is considered as the gold standard between MTB isolates.
method for the study of the epidemiology of tuberculosis, Finally, as Dr. Kreiswirth and Dr. Carrière, I would like
and its standardization has permitted to establish databases to speak about the new molecular marker, MIRU–VNTR.
allowing the comparison of the molecular typing results These markers are specific for MTB complex and they are
between laboratories. However, this technique requires a useful for molecular epidemiology studies of tuberculosis.
high amount of DNA (1 ␮g), and its protocol comprises A database comprising MIRU data for different MTB popu-
several steps contrary to PCR methods. It depends also on lations is also available. This molecular approach based on
the number of IS6110 copies, and for isolates with six or PCR is very reproducible, rapid, and shows a high power
less copies, the use of a secondary marker (spoligotyping, of discrimination in comparison with spoligotyping. More-
MIRU, PGRS . . . ) is indispensable for a better discrimina- over, this technique is a multilocus marker, thus, it can be
tion between strains. Moreover, RFLP-IS6110 data cannot used for the study of MTB population structure.
be used for population genetics study. Indeed for an haploid
organism as M. tuberculosis, the tests for studying the pop-
ulation structure are mainly based on the study of linkage 4. Fourth question: In your own TB molecular
disequilibrium (non-random association of genotypes oc- epidemiology approach, what is the specific
curring at different loci). And this technique does not reveal contribution of evolutionary genetics concepts
the variability of independent genetic loci and then cannot (population genetics/linkage disquilibrium analysis;
be used to analyze linkage disequilibrium. phylogenetic analysis), if any?
Several other genotyping methods based on PCR, have
been developed. The spoligotyping, for example, has shown 4.1. Response from LT
its potentialities to type M. tuberculosis strains, and to iden-
tify W-Beijing Families. Besides its use as a molecular My Ph.D. work concerns the study of the molecular
marker for genotyping MTB isolates, the spoligotyping per- epidemiology of tuberculosis in Casablanca, the economic
 L. Tazi et al. / Infection, Genetics and Evolution 2 (2002) 153–158 157

capital of Morocco, my country of origin. Up to now, few time and that human migration facilitates dispersion of the
genetic studies have been done on Moroccan MTB isolates. pathogen (via both dormant and active infection).
Thus, this work represents the first evolutionary genetics Evolutionary genetic concepts have greatly enhanced and
study performed in Morocco. in some instances augmented molecular epidemiological ap-
Two molecular techniques (RAPD and MIRU) have been proaches. Much of our molecular epidemiology is based on
used in this study in order to analyze the population struc- clonal theory, that is sister cells that share in common traits of
ture of the Moroccan population and to understand the dy- the parent. Furthermore, population genetics would suggest
namics of TB transmission in this country. The results ob- that certain medically relevant traits are non-randomly dis-
tained with the two markers have shown a significant linkage tributed along clonal or phylogenetic lines. That is, some lin-
disequilibrium, which represents evidence for clonal pop- eages of tuberculosis may possess specific traits that enable
ulation structure. Moreover, the population genetics tests successful host colonization (by evading the host immune
with phylogenetic studies have permitted to better under- response), pathogenesis, and efficient transmission (given by
stand TB burden in Morocco, and particularly in Casablanca. a clone-specific reproductive number). These concepts, in
As an example from these results, tuberculosis seems to conjunction with molecular biology, microbiology and epi-
have an ancient origin and an endemic situation in this demiology were used in an (ongoing) investigation of large
country. phylogenetic groups of M. tuberculosis strains as outlined
below.
In the early 1990s, my laboratory genetically character-
4.2. Response from CC
ized and monitored the spread of the highly drug-resistant
W strain which caused repeated “institutional” outbreaks in
The data provided by the molecular epidemiology ap-
both prisons and hospitals in New York (Bifani et al., 1996).
proach of the variability among natural population of M. tu-
The fact that nearly 85% of the infected population were
berculosis contributes to understanding their evolution and
HIV positive led to the early progression of disease and the
pathogenesis. The most important, but difficult to define
blossoming of case clusters. More than 500 W strain cases
parameter of a genotyping system is the time period over
have been identified in New York during the last decade and
which the degree of relatedness demonstrated is informa-
the isolates are commonly resistant to INH, RIF, STP, EMB,
tive. For example, it has been related that the evolution
PZA. The molecular analysis of these infecting isolates re-
rate of MIRU–VNTRs could be slightly slower than that of
vealed that the IS6110 pattern has evolved during the strains
IS6110 RFLP (Mazars et al., 2001). These observations sug-
transmission pathway among a New York population, and
gested that MIRU–VNTRs typing could be more appropri-
we have now identified nine closely related hybridization
ate than IS6110 RFLP for long-term epidemiological stud-
patterns. Although their IS6110 pattern vary, these strains
ies. In terms of phylogeny, the genetic relationship among
are confidently linked to the W strain outbreak on the ba-
M. tuberculosis isolates can be analyzed on dendrograms
sis of the identical array of DNA sequence mutations in the
obtained with a software-assisted analysis of patterns. This
numerous drug-resistant gene targets, including rpsL, katG,
can be done with the RFLP patterns as well as with the
rpoB, embB and pncA.
spolygotyping and MIRU–VNTR patterns.
The development of numerous genotyping methods, in-
I would like to point out that, as we develop more and
cluding spoligotyping and IS6110 insertion site mapping has
more refined molecular methods for analysis of the bacterial
identified additional molecular markers to link the W strain
genome do not we risk 1 day to consider as “epidemiologi-
to a very large phylogenetic lineage that is found in princi-
cally unrelated” drug-resistant bacteria which might only be
pal genetic group 1. Specifically, a deletion in the DR region
variants of the same clonal type?
and an IS6110 insertion in the origin of replication group ge-
For that purpose the molecular epidemiologist will need
ographically disparate isolates to this phylogenetic lineage
to interface with clinicians, epidemiologists and computer
(Kurepina et al., 1998). The current TB literature repeatedly
scientists to study for example an epidemic problem and
shows that members of this lineage, called the W-Beijing
more generally to interpret the results he obtained in term
strain family, successfully spread and cause disease in the
of evolution and phylogeny.
human population. Strain 210, a member of the W-Beijing
strain family and one that caused multiple outbreaks in west-
4.3. Response from BK ern states in the US is being sequenced at TIGR and its
genome will add to the data to unravel the genetic basis for
The molecular epidemiology of tuberculosis is a young the success of strains in this lineage (Bifani et al., 2002).
science that has grown with the advances in secondary The molecular epidemiology of the W-Beijing strain fam-
genotyping methods and with the understanding that strain ily in human populations indicate it has been successful, we
collections must be representative of a given population. In- now need to incorporate this information in other biolog-
terpreting the molecular data and the clustering of strains is ical systems such as in making decisions about strains in
complicated by fact tuberculosis is predominantly a chronic animal studies and in the choice of strains for vaccine chal-
reactive disease, that is most cases are reactive over a life- lenges. Therefore, the application or interpretation of evolu-
158 L. Tazi et al. / Infection, Genetics and Evolution 2 (2002) 153–158

tionary concepts in describing the molecular epidemiology Sreevatsan, S., Pan, X., Stockbauer, K.E., Connell, N.D., Kreiswirth,
(or disease burden) should be done with caution. Evolution- B.N., Whittam, T.S., Musser, J.M., 1997. Restricted structural gene
polymorphism in the Mycobacterium tuberculosis complex indicates
ary clock-speeds at certain genetic loci and with specific ge-
evolutionary recent global dissemination. Proc. Natl. Acad. Sci. U.S.A.
netic backgrounds are not well-understood or characterized 94, 9869–9874.
therefore, a multidisciplinary approach is recommended.
It should be emphasized that this data has been synthe-
sized over a decade and its growth is truly the result of a
collective group of laboratories around the world that have Loubna Tazi is a Moroccan PhD student at
shared their genotyping data through the internet, at meet- the Genetics of Infectious Diseases Laboratory
(UMR CNRS-IRD 9926) at the “Institut de
ings, and in collaborative studies. The result is that tuber-
Recherche pour le Développement” (IRD), Mont-
culosis is the model disease to study principals in bacterial pellier, France. Her thesis focuses on molecu-
molecular epidemiology. lar epidemiology of tuberculosis in Morocco, her
country of origin. This work is under the super-
4.4. MT vision of Dr. Anne-Laure Bañuls and Dr. Michel
Tibayrenc.
Since TB represents a major public health in her
This ends this very fruitful e-debate on a hot topic. I thank country, she is interested in studying the genetic diversity of M. tubercu-
you all three very much for participating in it. losis isolates circulating in the economic capital of Morocco, Casablanca.
This work will lead to determine the population structure of M. tuberculo-
sis and the dynamics of its transmission in this high incidence community.
References Loubna Tazi has participated to the students roundtable at the MEEGID
VI Congress in Paris, France (July 2002), and she has received the award
of best communication by a scientist from a developing country.
Bauer, J., Thomsen, V.O., Poulsen, S., Andersen, A.B., 1997.
False-positive results from cultures of Mycobacterium tuberculosis due
to laboratory cross-contamination confirmed by restriction fragment
length polymorphism. J. Clin. Microbiol. 35, 988–991.
Bifani, P.J., Plikaytis, B.B., Kapur, V., Stockbauer, K., Pan, X., Barry Kreiswirth is American. He was trained
Lutfey, M.L., Moghazeh, S.L., Eisner, W., Daniel, T.M., Kaplan, as a molecular biologist under the direction of Dr.
M.H., Crawford, J.T., Musser, J.M., Kreiswirth, B.N., 1996. Origin Richard Novick at the Public Health Research In-
and interstate spread of a New York City multidrug-resistant stitute. As a student, the focus of his research was
Mycobacterium tuberculosis clone family. JAMA 275, 452–457. the cloning and characterization of the staphylo-
Bifani, P.J., Mathema, B., Kurepina, N.E., Kreiswirth, B.N., 2002. Global coccal toxic shock syndrome toxin-1. The use of
dissemination of the Mycobacterium tuberculosis W-Beijing family molecular probes to identify the iatrogenic spread
strains. Trends Microbiol. 10, 45–52. of TSST-1 from a neurosurgeon to four patients
Fleischmann, R.D., Alland, D., Eisen, J.A., Carpenter, L., White, O., led to his interest in molecular epidemiology and
Peterson, J., DeBoy, R., Dodson, R., Gwinn, M., Haft, D., Hickey, the tracking of nosocomial infections.
E., Kolonay, J.F., Nelson, W.C., Umayam, L.A., Ermolaeva, M., The focus of his laboratory is to develop the use of objective and rapid
Salzberg, S.L., Delcher, A., Utterback, T., Weidman, J., Khouri, H., molecular targets to control the spread of both nosocomial infections, es-
Gill, J., Mikula, A., Bishai, W., Jacobs Jr., W.R., Venter, J.C., Fraser, pecially methicillin resistant Staphylococcus aureus, and the global spread
C.M., 2002. Whole-genome comparison of Mycobacterium tuberculosis of M. tuberculosis. In this regard they have used DNA sequence compar-
clinical and laboratory strains. J. Bacteriol. 184, 5479–5490. ison of variable number tandem repeat sequences in the S. aureus protein
Gascoyne-Binzi, D.M., Barlow, R.E.L., Frothingham, R., Robinson, G., A gene as a model to rapidly identify and track this pathogen in both the
Collyns, T.A., Gelletlie, R., Hawkey, P.M., 2001. Rapid identification hospital and the community. Their current goal is to develop a battery of
of laboratory contamination with Mycobacterium tuberculosis using DNA sequence targets that will provide an objective approach to build
variable number tandem repeat analysis. J. Clin. Microbiol. 39, 69–74. large relational databases to control the spread of bacterial pathogens.
Kurepina, N.E., Sreevatsan, S., Plikaytis, B.B., Bifani, P.J., Connell, N.D.,
Donnelly, R.J., van Sooligen, D., Musser, J.M., Kreiswirth, B.N., 1998.
Characterization of the phylogenetic distribution and chromosomal
insertion sites of five IS6110 elements in Mycobacterium tuberculosis:
non-random integration in the dnaA–dnaN region. Tuber. Lung Dis.
79, 31–42. Christian Carrière, born in Montpellier, France,
Mazars, E., Lesjean, S., Bañuls, A.L., Gilbert, M., Vincent, V., Gicquel, is French. He is head of the bacteriology service
B., Tibayrenc, M., Locht, C., Supply, P., 2001. High-resolution VNTR of the Montpellier Hospital. His lab has an activity
typing as a portable approach to global analysis of Mycobacterium of clinical diagnosis in bacteriology and particu-
tuberculosis molecular epidemiology. Proc. Nat. Acad. Sci. U.S.A. 98, larly in mycobacteriology. Besides his activity of
1901–1906. clinical microbiologist he has performed research
Ramose, M.deC., Soini, H., Roscanni, G.C., Jaques, M., Villares, M.C., on molecular epidemiology of nosocomial infec-
Musser, J.M., 1999. Extensive cross-contamination of specimens tions and has worked in the determination of an-
with Mycobacterium tuberculosis in a reference laboratory. J. Clin. tibiotic susceptibilities of M. tuberculosis strains
Microbiol. 37, 916–919. by using luciferase reporter mycobacteriophages.