biosensors

Review
Recent Advances in Surface Plasmon Resonance Sensors for
Sensitive Optical Detection of Pathogens
Joon-Ha Park, Yeon-Woo Cho and Tae-Hyung Kim *

School of Integrative Engineering, Chung-Ang University, 84 Heukseuk-ro, Dongjak-gu, Seoul 06974, Korea;
joonha95@cau.ac.kr (J.-H.P.); tree7391@cau.ac.kr (Y.-W.C.)
* Correspondence: thkim0512@cau.ac.kr

Abstract: The advancement of science and technology has led to the recent development of highly
sensitive pathogen biosensing techniques. The effective treatment of pathogen infections requires
sensing technologies to not only be sensitive but also render results in real-time. This review thus
summarises the recent advances in optical surface plasmon resonance (SPR) sensor technology, which
possesses the aforementioned advantages. Specifically, this technology allows for the detection of
specific pathogens by applying nano-sized materials. This review focuses on various nanomaterials
that are used to ensure the performance and high selectivity of SPR sensors. This review will
undoubtedly accelerate the development of optical biosensing technology, thus allowing for real-time
diagnosis and the timely delivery of appropriate treatments as well as preventing the spread of highly
contagious pathogens.

Keywords: SPR; optical sensor; graphene oxide; gold; nucleic acid; biosensing techniques

1. Introduction



Pathogens are defined as foreign antigens that have adverse effects, such as (1) pro-


ducing toxins, (2) penetrating tissues, (3) colonising tissues, (4) intercepting nutrients,
Citation: Park, J.-H.; Cho, Y.-W.; Kim, and (5) immunosuppressing the host [1–3]. Particularly, contagious pathogens (e.g., those
T.-H. Recent Advances in Surface
that cause bacterial urinary tract infection, malaria, influenza virus, dengue virus, human
Plasmon Resonance Sensors for
immunodeficiency virus (HIV), and severe acute respiratory syndrome coronavirus (SARS-
Sensitive Optical Detection of
CoV-2) are capable of human-to-human presymptomatic transmission [4–9]. Infection
Pathogens. Biosensors 2022, 12, 180.
pathways with receptor-ligand interactions (RLIs) constitute a representative mechanism
https://doi.org/10.3390/bios12030180
through which pathogens bind to their target [10–13]. Additionally, given that RLIs are
Received: 10 February 2022 biological events that occur in living cells, a high-sensitivity diagnostic sensor is required
Accepted: 11 March 2022 to detect pathogens [14]. Therefore, biosensing approaches must be constantly optimised
Published: 17 March 2022 to achieve better signal sensitivity.
Publisher’s Note: MDPI stays neutral To increase the sensitivity and selectivity between specific pathogens, recent studies
with regard to jurisdictional claims in have developed and evaluated novel diagnostic tools that incorporate pathogen binding
published maps and institutional affil- mechanisms and antigen capture strategies, such as antigen–antibody and aptamer–ligand
iations. interactions [15,16]. Current diagnostic procedures are based on several clinicopathological
analysis methods, such as gene sequencing, polymerase chain reaction (PCR), fluorescence
methods, mass spectrometry, and enzyme-linked immunosorbent assays (ELISA) [17–24].
However, although these bioanalytical methods are highly accurate, each of them has
Copyright: © 2022 by the authors. unique limitations. For example, (1) the fluorescence method requires labelling the sample
Licensee MDPI, Basel, Switzerland. with a fluorescent probe, which adds complexity to the procedure; (2) the analysis of
This article is an open access article multiple samples using mass spectrometry is highly time-consuming; and (3) most of the
distributed under the terms and
aforementioned techniques are generally expensive and require sophisticated equipment
conditions of the Creative Commons
and specialised processing, in addition to having a high response time [25–30]. To ad-
Attribution (CC BY) license (https://
dress these limitations, bioanalytical techniques based on optical biosensors have recently
creativecommons.org/licenses/by/
garnered increasing attention [31].
4.0/).

Biosensors 2022, 12, 180. https://doi.org/10.3390/bios12030180 https://www.mdpi.com/journal/biosensors

Biosensors 2022, 12, 180 2 of 17

Particularly, plasmonic-based biosensors constitute a next-generation technology that

facilitates the miniaturisation of biosensors, thereby enhancing detection throughput and
reducing operating costs [32]. Pathogen detection can be based on a variety of plas-
monic phenomena, including SPR, localized SPR, surface-enhanced Raman scattering,
surface-enhanced infrared absorption spectroscopy, and surface-enhanced fluorescence [33].
Among these novel approaches, surface plasmon resonance (SPR)-based sensing techniques
are a promising new means to detect pathogens quickly with high sensitivity, and therefore,
this method has begun to replace other conventional diagnostic approaches. SPR is an
electromagnetic surface wave that propagates parallel to the interface region. On the basis
of this principle, changes in a given sample can be detected by monitoring the intensity
of the SPR angle (θ) reflected after the plasmon wave of a medium with a low refractive
index permeates [34,35]. SPR sensors have some minor disadvantages, such as the possi-
bility of sample SPR peak deflection when a strong electromagnetic field acts on sensitive
biological samples [36]. However, there are four main advantages that compensate for
these drawbacks: (1) this approach is non-destructive and non-invasive—in other words,
it does not cause any damage or physicochemical transformation to the sample; (2) it
allows for the sensitive optical quantification of the interaction between molecules in real-
time without the need for a separate probe using a radioactive or fluorescent material;
(3) after the detection of the target analyte, the surface of the sensor can be reactivated
and used repeatedly upon treatment with a regeneration solution; and (4) the lifetimes of
nanoparticles can be flexibly controlled and extended through surfactant surface coating or
chemical conjugation (e.g., hybrid nanomaterials) [37–41]. Additionally, this approach is
rapid, accurate, cost-effective, requires no specialised training, and can be applied to detect
a wide variety of targets [42–44]. Collectively, these advantages make SPR-based sensing
techniques especially well-suited for real-time diagnosis both in clinical laboratories and in
the field [45–47].
SPR sensors based on novel materials are actively being developed. For example,
some metals, such as gold film or particle-based SPR platforms, are used to facilitate particle
generation and optimise plasmonic performance [48–55]. For instance, graphene-based SPR
platforms have excellent analyte adsorption properties, great corrosion resistance, and high
thermal resistance [56–59]. Hybrid graphene/gold-based SPR platforms maximise plas-
monic performance via graphene–gold interactions [60–64]. Aptamer-based SPR sensors
reportedly exhibit uniquely high analytical specificity [65–68]. Three-dimensional (3D)
structure-based SPR platforms possess a higher surface area than their 2D counterparts,
which increases the likelihood of the analyte binding to the platform [69–71]. Additionally,
these nanomaterials are not only highly biocompatible but also have fewer negative envi-
ronmental impacts. In fact, some of these materials are actively used for environmental
remediation [72,73]. SPR biosensors are therefore often used as a preliminary diagnostic
tool, as they enable efficient and high-throughput diagnosis due to their fast detection
time and the fact that they do not require labelling [74,75]. In this section, SPR platforms
were classified into five categories depending on the materials used for pathogen sensing:
(1) graphene oxide-based, (2) gold-based, (3) hybrid graphene/gold-based, (4) nucleic
acid-based, and (5) 3D structure-based (Table 1). In this review, different SPR-based sensing
techniques that can rapidly detect pathogens, such as viruses and bacteria, are investigated
and discussed (Figure 1).
Biosensors 2022, 12, 180 3 of 17

Table 1. Materials used for the construction of SPR sensors for the optical detection of pathogens.

Materials Pathogen/Disease LOD Ref.

Graphene oxide film Leptospirosis - [76]
BaTiO3 -adsorbed graphene oxide Pseudomonas 220 deg/RIU [77]
Cadmium sulphide quantum dot-adsorbed
Dengue virus E-protein 0.001 nM [78]
graphene oxide
Gold film SARS-CoV-2 1.02 pM [79]
Antibody-conjugated gold nanoparticles Campylobacter jejuni 8 × 106 CFU/mL [80]
Gold nanospikes SARS-CoV-2 0.5 pM [81]
Gold–aluminium bilayer DENV2-NS1 0.1 µg/mL [82]
Hybrid graphene/gold film Dengue virus 28 fM [83]
Spherical gold nanoparticles on graphene A and O-type foot-and-mouth A-types: 100 fg/mL
[84]
oxide film disease virus O-types: 100 pg/mL
Graphene-encapsulated gold nanoparticles Hepatitis B 0.05 pg/mL [85]
Aptamer on gold film Shigella sonnei - [86]
Aptamer–aptamer sandwich formation Vibrio fischeri 103 –104 CFU/mL [87]
Sandwich formation with gold nanorods Norovirus capsid protein 70 aM [88]
Gold nanowire-type Epstein-Barr virus 4.1 × 10−5 RIU [89]
Gold nanoprisms Rotavirus 126 ± 3 PFU/mL [90]
MoS2 -coated gold optical fibre Escherichia coli 94 CFU/mL [91]

Figure 1. Schematic of recent advances in SPR sensor technology with various nanomaterials and
applications for pathogen detection.
Biosensors 2022, 12, 180 4 of 17

2. Graphene Oxide-Based SPR Sensor

The edge of the graphene oxide (GO) structure is rich in oxygen functional groups;
it is a single-layer, two-dimensional carbon material [92,93]. GO has outstanding properties
that show great potential for biosensing at the molecular level because of its abundant
π-conjugation structure along with its large surface-to-volume ratio [94]. Therefore, GO
has specific intrinsic features that enable the robust immobilization of biomolecules on the
SPR biosensor platform.
A recent study reported the detection of Leptospirosis bacteria using a graphene-
based SPR sensor [76]. The researchers coated a glass platform with graphene to effectively
adsorb the bacteria. Rodent urea was then used as the main sample, and the analytical
outcomes were classified as polyuria (high urine production) and oliguria (low urine
production). Upon measuring the urea with the SPR platform, polyuria was defined be-
tween 128.8378 (deg/RIU) and 130.40 (deg/RIU), whereas oliguria was defined between
134.5118 (deg/RIU) and 131.0978 (deg/RIU) with different angles of reflection (Figure 2a).
The proposed graphene-based SPR sensor has been theoretically and numerically anal-
ysed to detect Leptospirosis-causing bacteria in rodent urine, suggesting that it can be
utilised for the early detection of this disease with high sensitivity and accuracy (Figure 2b).
Although graphene can be used to increase SPR signals, additional efforts have been made
to improve sensitivity and detection accuracy. To achieve this, researchers have begun to
study barium titanate nanocomposites. Through the adsorption of barium titanate onto
graphene, the proposed sensor achieved a higher detection sensitivity compared to a single
graphene-based SPR biosensor. Furthermore, Z. Xia et al. proposed an SPR sensor based
on barium titanate and also achieved high analyte sensitivity [95]. Another study detected
Pseudomonas bacteria by mixing graphene and barium titanate [77]. The platform was
fabricated by deposition of silver and graphene on a BK7 prism glass plate that refracts in-
cident light, and the sensors were constructed in the presence or absence of barium titanate
(BaTiO3 ) nanocomposites. The authors reported that the maximum sensitivity, detection
accuracy, and quality parameters of the sensor were 220 deg/RIU, 7.09, and 101.38 RIU−1 ,
respectively. Therefore, this SPR platform can sensitively detect Pseudomonas bacteria.
Additionally, the authors demonstrated that the graphene thickness had a critical effect on
the results even when the affinity of the layers changed, and the monolayer exhibited the
best reflectivity curve.
Quantum dots are a nanomaterial with a high surface area and signal-to-noise ratio,
and significantly improved data accuracy [96]. Additionally, combining graphene oxide
with carboxyl functional groups and quantum dots greatly increases mechanical stability
as well as the potential for SPR signal amplification [97]. A study from 2019 reported the
detection of dengue virus E-protein by adsorbing cadmium sulphide quantum dots on
an optical SPR platform based on graphene oxide [78]. The SPR platform with cadmium
sulphide quantum dots was fabricated through the stirring method, and the matrix of the
sensor was based on graphene oxide. The fabricated SPR platform achieved a detection
limit of 0.001 nM for the dengue virus E-protein, which demonstrated its high sensitivity,
and the estimable linear range of the sensor was 0.0001 to 0.01 nM. These sensing results
suggest that graphene oxide can improve the performance of SPR biosensors and is thus a
promising candidate in the field of biosensing.
The above-described observations demonstrate that graphene oxide-based materials
have various advantages for pathogen detection. It has been proven that GO does not dam-
age biological samples, nor does it interfere with the SPR effect. Moreover, because of the
characteristics of GO, such as its easy handling and excellent analyte adsorption properties,
highly sensitive GO-based SPR sensors are expected to be developed in the future.
Biosensors 2022, 12, 180 5 of 17

Figure 2. (a) SPR signal intensity vs. incident angle plot with polyuria (left) and oliguria (right).
(b) Variation in parameters for different sensing medium refractive indices in pure water, polyuria,
and oliguria. With permission from [76], Copyright 2020, Springer.

3. Gold-Based SPR Sensor

Gold-based materials can be easily synthesised and are highly biocompatible. Addi-
tionally, these materials are structurally strong, and their optical properties can be fine-
tuned depending on the conditions of the platform. Therefore, gold is widely used in
biosensing chips [98,99]. Over the past decade, gold has been applied in various forms,
and previous studies have demonstrated that this material exhibits strong red-shifting
properties [100]. Furthermore, the strong radioactive properties of gold particles, such as ab-
sorption, scattering, and plasmonic field generation, can be widely applied to SPR biochips.
Bong et al. developed a gold-based SPR sensor for SARS-CoV-2 [79]. The sensor was
fabricated by binding antibodies through chemical conjugation on the gold chip sensor,
which was the base of this SPR platform. The limit of detection for an analyte isolated from
serum was 1.02 pM (i.e., the sensor was highly sensitive). Additionally, the sensor only
detected coronavirus and no other influenza viruses, thus demonstrating its high selectivity
for the analyte due to the characteristics of the substrate. Moreover, the gold-based SPR
sensor rendered results very quickly, making this approach uniquely well-suited for the
accurate detection of diseases at an early stage.
Spherical gold nanoparticles have various advantages compared to gold films or
random gold particles, such as their narrow size distribution, consistent shape and size,
and easy handling. Particularly, the size specificity of gold nanoparticles results in a narrow
standard deviation from the analyte signal during SPR measurement. Furthermore, anti-
bodies that specifically recognise various chemical conjugates or analytes can be attached
to the spherical particles, thereby taking advantage of the large surface area of the nanopar-
ticles and enhancing the SPR signal. A study from 2017 reported the development of a
surface plasmon sensor using an antibody on a gold-based platform to sense pathogenic
bacteria [80]. The fabricated biosensor was based on the principle of strengthening the
surface plasmon effect by adding gold nanoparticles (AuNPs) after binding the analyte on
the gold sensor chip. The limit of detection for the analytes of this fabricated SPR platform
was 8 × 106 CFU/mL, and the sensor was specifically sensitive to the pathogenic bacterium
Campylobacter jejuni. Furthermore, the AuNPs in the sensor increased the efficiency of
Biosensors 2022, 12, 180 6 of 17

analyte analysis, whereas the gold sensor chip enhanced SPR sensitivity. Interestingly,
although the use of AuNPs enhanced SPR performance, the SPR effect was maximised
when using other types of gold nanostructures, such as nano-urchins or nanospikes.
Similar to sea urchins, gold nanospikes are not perfectly circular but have a sharp and
uneven surface. This unique structure could enhance the red-shifting of the surface plasmon
resonance absorption peak, in addition to increasing the electromagnetic field at the tip of
the spike structure compared to that of a spherical particle. A recent study developed a
gold spike nanoparticle-based SPR sensor that detects SARS-CoV-2 in human plasma [81].
The researchers deposited the gold spike nanoparticles through electrodeposition onto a
glass slide rather than gold in its bulk state (Figure 3a). The fabricated SPR sensor exhibited
a low limit of detection of 0.08 ng/mL (~0.5 pM) for SARS-CoV-2 in addition to being highly
specific, as it did not react to three competing substances and BSA protein (Figure 3b,c).
This shows that the fabricated SPR sensor could be used for the early diagnosis of COVID-19
and can accurately recognise virus analytes without detecting other substances remaining
in the human plasma.

Figure 3. (a) Schematic illustration of the fabricated gold nanospike-based SPR sensor. (b) Plot
representing the detection limit for the analyte. (c) Selective affinity test against other interference
substances. With permission from [81], Copyright 2020, ELSEVIER.

Studies have demonstrated that bimetallic combinations can provide greater sensitivity,
signal-to-noise ratio (SNR), and increased operating range values, all of which cannot be
achieved with a single metal layer. A recent study sought to offset the shortcomings
of single metal layer-based sensors and improve the performance of the SPR sensor by
using a novel bimetal layer structure of Au and aluminium (Al) [82]. The researchers
detected the DENV2 NS1 protein with the developed SPR sensor by laying a gold and
aluminium bilayer on the SU-8 spacer. The fabricated SPR sensor recorded a limit of
detection of 0.1 µg/mL for DENV2-NS1, and the linear range of the sensor was between
0.1 and 10 µg/mL. Interestingly, the limits of detection in both PBS and blood plasma
were also within a similar range (i.e., as low as 0.1 µg/mL), suggesting that this approach
can detect viruses directly in blood samples or liquid biopsies. This study demonstrated
that a specific virus can be detected with high sensitivity through immediate screening in
clinical samples.
According to a recent study, on-chip integration of electronics and plasmonics would
allow for samples to be guided to relevant locations and sensed thereafter [101–104].
Particularly, gold-based plasmonic trapping technology could substantially improve the
efficiency of broadband photodetectors and SPR sensors [105,106]. Gold nanostructures
have a virtually infinite potential depending on how they are applied. These structures
can be constructed using various methods, such as growing gold on a substrate, attaching
it by chemical conjugation, or depositing it. Similarly, the ability of the sensor to detect
the analyte can be fine-tuned. The implementation of gold in the SPR sensor enhances its
surface plasmon effect, the most basic characteristic of metal, and causes no damage to the
platform body, both of which are considered crucial advantages of this material.
Biosensors 2022, 12, 180 7 of 17

4. Hybrid Graphene/Gold-Based SPR Sensor

Gold-based biosensors are currently preferred over other sensors based on other metals.
However, according to a recent study, there is a risk of peeling if the gold layer coating on
the platform surface becomes too thin [107]. Furthermore, another study reported that gold
itself has little influence on the adsorption of biomolecules, which can lower the detection
limit [108]. Additionally, sensors based only on graphene exhibit an excellent adsorption
force for the detection of analytes, but their surface plasmonic effect is significantly lower
than that of metal-based sensors [109]. To compensate for this limitation, several recent
studies have developed biosensors based on a hybrid graphene/gold structure.
Omar, N.A.S. et al. developed a hybrid graphene/gold sensor to detect the dengue
virus by thinly coating reduced graphene on a gold film [83]. The generated SPR plat-
form was coated with gold in the form of a film through deposition on a glass substrate,
and graphene was thinly coated through spin-coating (Figure 4a). This hybrid sensor
achieved a detection limit of 28 fM and a linear range of 0 pM to 10 pM, as determined
with a dengue virus solution (Figure 4b). A sample of 100 pM human serum albumin
(HSA) was used for the selectivity test, and the fabricated platform reacted only with the
dengue virus and showed low reactivity to HSA (Figure 4c). These results demonstrated
that the hybrid graphene/gold SPR sensor could potentially be used in health and envi-
ronmental monitoring. In addition to gold films, gold nanoparticles, an important type
of plasmon nanoparticles, possess excellent plasmonic properties and have thus begun
to be implemented in a variety of applications. The use of particles maximises the sur-
face area of the gold film and graphene oxide complex. At the same time, the spherical
gold nanoparticles adsorb to the graphene to enhance adsorption and plasmonic perfor-
mance [84]. An SPR sensor was developed by combining graphene oxide and spherical
gold nanoparticles through sonication and stirring. The researchers detected A-type and
O-type foot-and-mouth disease viral particles through the resulting substrate and achieved
limits of detection of 100 fg/mL and 100 pg/mL, respectively. Because of the characteristics
of the GO on the substrate, single-stranded DNA (ssDNA) can readily bind to the sensor
through electrostatic interactions, and thus the limit of detection is 1000 times lower than
that of conventional PCR. Therefore, combining pristine gold nanoparticles and GO can
greatly improve the limit of detection of biosensors.
Many SPR sensors have been developed to detect analytes by adsorbing gold nanopar-
ticles on graphene; however, this method is notorious for its limitations. Specifically,
a recent study reported that the performance of the sensor was low because the adsorbed
gold nanoparticles tend to agglomerate on the platform surface, and the absorbance curve
is widened, thereby decreasing sensor stability [110]. To solve this problem, recent stud-
ies have explored the encapsulation of gold nanoparticles using graphene. For example,
a related study used GO to encapsulate gold nanorods, thereby overcoming the aforemen-
tioned limitations [85]. Concretely, the researchers developed a GO-encapsulated gold
nanorod-based SPR sensor to detect hepatitis B, and this platform was fabricated through
sulphonylation. To increase the specificity of the sensor towards hepatitis B, HBsAg an-
tibodies were decorated on the GO surface. The limit of detection for hepatitis B was
0.05 pg/mL, and the sensor had a wide linear range of 1–1000 pg/mL. The researchers
then diagnosed HBV-infected patients with the fabricated platform and achieved a relative
standard deviation (RSD) of less than 5%, thus demonstrating the potential applicability of
the sensor in clinical diagnosis.
Biosensors 2022, 12, 180 8 of 17

Figure 4. (a) Schematic illustration of the biosensor fabrication process. (b) Plot indicating the limit
of detection and linear range. (c) Selectivity of the fabricated platform. With permission from [83],
Copyright 2019, MDPI.

A hybrid platform decorated with a gold film or nanoparticles on graphene oxide

showed the potential to enhance the surface plasmon resonance performance of the sensor.
Additionally, by taking advantage of the properties of GO, an antibody that can specifically
recognise an analyte can be adsorbed on the surface, thus allowing for the biosensor to be
used alongside immunoassays. This type of sensor is much more versatile because it com-
bines the advantages of GO and gold. Additionally, sensing performance is substantially
higher than that of sensors based on GO or gold alone. Nevertheless, additional studies are
required to better understand this hybrid technique and which factors can be modified to
optimise its performance.

5. Nucleic Acid-Based SPR Sensor

Aptamer-based biosensors have recently garnered increasing attention because they
integrate the advantages of antibody-based sensors while also exhibiting excellent thermal
stability, low cost, and an extremely wide variety of applications [111–113]. Furthermore,
unlike antibodies, nucleic acid ligand aptamers can be chemically synthesised and easily
modified using functional groups, in addition to being highly stable [114]. When this
biomaterial binds to an analyte of interest, it has a similar or stronger binding force than
that of antibodies, thus enabling stable sensitivity measurements [115]. Because of these
characteristics, aptamers can be widely applied in the fields of biosensors, diagnostics,
and research agents.
Biosensors 2022, 12, 180 9 of 17

A study conducted in 2018 reported the detection of virulence factors using an aptamer-
based SPR sensor [86]. The SPR sensor was fabricated by binding the target protein on
a gold film through various chemical conjugation steps. The aptamer was selected to be
specific to the analyte through the systematic evolution of ligands via the exponential
enrichment (SELEX) technique. The sensor exhibited a low limit of detection for Shigella
sonnei, and the measurable linear range was 0–100 nm/mL. Except for S. sonnei, all other
Shigella and non-Shigella species exhibited a low recognition rate, thus demonstrating the
high specificity of the sensor. This is an example of aptamer selection through SELEX, which
substantially enhanced the analyte selectivity for a specific pathogen species. Specifically,
the incorporation of aptamers dramatically increased the selectivity to the analyte, but the
approach was not sufficiently sensitive. To solve this problem, the authors proposed a
method to enhance sensitivity by attaching a pair of aptamers to a specific analyte.
A sensor with a pair of aptamers in a sandwich configuration was reported to improve
the sensitivity and specificity on various platforms. Therefore, this approach has recently
been implemented to optimise SPR biosensors. The use of a secondary aptamer amplifies
the signal, and these types of biosensors are suitable for fast diagnosis because they allow
for the real-time monitoring of any signal changes. A recent study employed this sandwich
aptamer technique to construct an SPR sensor to detect pathogens [87]. The fabricated
SPR platform consisted of an aptamer attached to a paper strip sensor. The sensor became
red upon contact with the analyte, and the colour shift could be quantified using the SPR
principle. The limit of detection for the analyte was 103 –104 CFU/mL, and the sensor had
a useful analytical range of 4 × 101 –4 × 105 CFU/mL. Moreover, because of its aptamer
properties, the sensor exhibited low specificity for other analytes, except for Vibrio fischeri.
The authors thus concluded that this paper strip-based sensor could be used for the real-
time detection of analytes in the field.
Although the sandwich aptamer technique can achieve high levels of sensitivity,
its plasmonic effect could be further improved. Therefore, studies on the binding of metals
to the secondary aptamer have been actively conducted, and a recent study demonstrated
that the SPR effect could be improved by binding gold nanoparticles [88]. The researchers
connected the primary aptamer to a gold chip through chemical conjugation, after which the
norovirus capsid protein was bound (Figure 5a). The secondary aptamer was bound to the
analyte while attached to gold nanorods, which were fabricated through the seed-growth
method. The limit of detection of the generated SPR sensor was approximately 70 aM,
thus demonstrating that the sensor was very sensitive to the analyte (Figure 5b). Notably,
the signal was 105 times stronger than when gold nanorods were not used (Figure 5c).
Additionally, by using two aptamers as capturing reagents for the norovirus capsid protein,
non-specific binding events of other substances could be minimised. The aptamer–aptamer
surface sandwich SPR platform with nanorod amplification is a promising means for the
direct analysis of difficult-to-detect virus samples, and a wider variety of analyte-specific
aptamers is expected to be developed in the future.
Prior to the application of aptamers to the SPR sensor, this technology developed
rather slowly. However, aptamer-based sensors have been stably developed over the past
decade to this day. Aptamers are commonly selected by SELEX, but this technique is known
for its limitations in targeting pathogens. This is because the highly variable and complex
structure of the pathogen can affect the performance of the aptamer. Therefore, a simpler
and more efficient SELEX method must be developed to facilitate the synthesis of universal
aptamers for various pathogens. Aptamer–analyte capturing methods have already been
developed, and the limitations of existing aptamers can almost be fully overcome by
binding metals to the secondary aptamer, among other strategies. Nevertheless, additional
efforts are needed to broaden the applicability of these promising sensing technologies
while also solving their limitations.
Biosensors 2022, 12, 180 10 of 17

Figure 5. (a) Schematic illustration of SPR sensor fabrication via the sandwich aptamer technique.
(b) Linear plot of norovirus spike protein-sensing performance. (c) Linear plot of norovirus spike
protein-sensing performance without gold nanorods. With permission from [88], Copyright 2018,
American Chemical Society.

6. 3D Structure-BASED SPR Sensor

Simple two-dimensional structures greatly limit the application of the plasmonic effect
for pathogen sensing. Additionally, the attachment of chemical conjugates, aptamers,
and antibodies significantly affects the surface area of the sensor, which has been a major
obstacle for platform development [116]. In response to these limitations, 3D structures
began to be implemented in biosensors, particularly to increase surface area. Moreover,
the three-dimensional nature of these structures enhances the electromagnetic field at the
interface between the structural gaps, which improves pathogen detection.
A recent study successfully detected the Epstein-Barr virus (EBV) using this 3D struc-
ture approach [89]. The SPR 3D structure sensor developed by the researchers was made
through nanoimprinting lithography, and a nanowire-type platform was constructed via
gold deposition. The limit of detection for the analyte was 4.1 × 10−5 RIU, and the sensor
exhibited high specificity. The developed 500 nm periodic nanowire 3D structure SPR
sensor could successfully detect DNA and genes without labelling. Furthermore, the de-
tection system developed in this study was not only cost-effective but also had excellent
measurement and production repeatability.
To further maximise the efficiency of the electromagnetic field between the 3D-shaped
structures, researchers have developed a platform in which certain shapes are tightly
packed. This approach not only increases the surface area further but also enhances the
SPR signal efficiency for pathogen detection. For example, a recent study reported the
detection of rotavirus via the application of an octupolar nanopattern to a platform [90].
The researchers formed gold nano prisms on the sensor with an electron-beam lithography
(EBL) system, which was based on octupolar geometry. The limit of detection of the
fabricated platform was 126 ± 3 PFU/mL, which was lower than the plaque-forming
unit (1 × 103 PFU/mL) of rotavirus in water, thus demonstrating a sensitive detection
Biosensors 2022, 12, 180 11 of 17

potential. To measure the selectivity of the generated platform, the researchers measured
bovine herpesvirus (BHV1) and equine viral arteritis (EVA), neither of which elicited a
response from the sensor. In this way, the disadvantages of the existing method could
be overcome, and current findings suggest that efficient pathogen detection can be cost-
effectively achieved using portable point-of-care approaches.
In recent years, many 3D optical fibre immunosensors based on surface modification
using nanomaterials have been reported to improve biocompatibility [117–119]. Addi-
tionally, SPR sensors combining 2D material nanosheets and 3D fibres have been recently
developed. Particularly, the properties of 2D MoS2 nanosheets have attracted considerable
interest from the scientific community because of their broad applicability in the field of
biosensing [120]. This combination has significant advantages: (1) high electron mobility,
(2) low toxicity, (3) high surface area, and (4) thermal stability. A recent study reported a 3D
fibre optic SPR sensor for quantitative analysis of E. coli (Figure 6a) [91]. An optical fibre
SPR sensor was developed through ultrasonication and etching, after which MoS2 was
coated on the cylindrical fibre surface (Figure 6b). The limit of detection of the fabricated
SPR platform was 94 CFU/mL (Figure 6c), and the sensor exhibited excellent selectivity for
E. coli because cross-reaction with other materials was prevented by the MoS2 coated on the
surface (Figure 6d). This 3D optic fibre SPR sensor exhibited significantly higher sensitivity
and in vivo functionalization compared to other similar sensors, in addition to rendering
rapid results. However, the instability of the antibody increased after a certain amount of
time, and defects in the surface or edge of the MoS2 nanosheets caused sulphur vacancies.
Therefore, more studies are needed to improve the stability and robustness of the sensor.

Figure 6. (a) Schematic illustration of an experimental setup to test sensors. (b) Field emission
scanning electron microscope (FESEM) image of the fabricated optic fibre surface. (c) Transmission
spectra plot indicating the limit of detection. (d) Atomic force microscope image analysis of the
fabricated optic fibre surface. With permission from [91], Copyright 2018, Elsevier.

Interestingly, the reported 3D structure-based SPR sensor exhibited a high surface

area, excellent electromagnetic field efficacy, and SPR signal enhancement ability due to its
material and structural properties. Particularly, the sensor was able to detect pathogens in
Biosensors 2022, 12, 180 12 of 17

real-time in the field with a lower limit of detection compared to conventional 2D materials
alone. It has been proven that the combination of 3D structures and 2D materials not only
enhances in vivo functionalization but can also maximise SPR-sensing performance. By tak-
ing advantage of fact that the results differ depending on how the structures and materials
are utilised, 3D-based sensors can be excellent candidates for the effective detection of a
wide variety of targets.

7. Conclusions
This review summarised recent research on optical SPR platforms for pathogen de-
tection via the application of different materials. All the different nanomaterials reported
above have been proven to exhibit excellent efficacy for pathogen detection. In the case
of aptamer-based SPR sensors, rather than simply performing analysis with one aptamer,
the sandwich aptamer technique was implemented, and gold nanorods were combined
with a secondary aptamer to improve the detection limit of difficult-to-analyse pathogens.
Platforms with a 3D nanostructure increased the effectiveness of the electromagnetic field
through the arrangement of uniform structures at a high density. The 3D nanostructure
led to a strong enhancement of the SPR signal, and these features allow for the rapid
detection of pathogens in the field. Therefore, each nanomaterial has its own unique optical
properties and various reaction patterns to analytes, and it is expected that the sensing
potential of SPR sensors can be maximised by offsetting their disadvantages through the
fusion of these materials or the synthesis of novel alternatives.
Increasing numbers of human diseases are reported each year. Therefore, researchers
need to develop a sensor that can easily detect different kinds of pathogens. In many
situations, the analytes of interest are present in low concentrations in complex media.
Therefore, additional efforts are required to improve the materials and technology on
which biosensors rely to improve their sensitivity, accuracy, and specificity. Furthermore,
the sensing performance of platforms with complex structures is often superior, but this
commonly reduces stability. Therefore, in addition to enhancing detection potential, efforts
must be made to improve or at least preserve the stability, robustness, and storability of
biosensors, as well as to avoid any potential problems when they are used in the field.

Author Contributions: Writing—original draft, J.-H.P.; Conceptualization, J.-H.P., Y.-W.C. and

T.-H.K.; resource and data curation, J.-H.P.; writing—correction and editing, Y.-W.C. and T.-H.K.
All authors have read and agreed to the published version of the manuscript.
Funding: This work was supported by the Chung-Ang University Research Scholarship Grants
in 2020 and by the National Research Foundation of Korea (NRF) grant funded by the Korean
government (MSIT) (No. 2019M3A9H2031820).
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: Not applicable.
Conflicts of Interest: The authors declare no conflict of interest.

References
1. Ingrao, F.; Rauw, F.; Lambrecht, B.; Van den Berg, T. Infectious Bursal Disease: A complex host–pathogen interaction. Dev. Comp.
Immunol. 2013, 41, 429–438. [CrossRef] [PubMed]
2. Beltran, P.J.; Federspiel, J.D.; Sheng, X.; Cristea, I.M. Proteomics and integrative omic approaches for understanding host–pathogen
interactions and infectious diseases. Mol. Syst. Biol. 2017, 13, 922. [CrossRef] [PubMed]
3. Mangen, M.-J.J.; Plass, D.; Havelaar, A.H.; Gibbons, C.L.; Cassini, A.; Mühlberger, N.; Van Lier, A.; Haagsma, J.A.; Brooke, R.J.;
Lai, T.; et al. Correction: The Pathogen- and Incidence-Based DALY Approach: An Appropriated Methodology for Estimating the
Burden of Infectious Diseases. PLoS ONE 2013, 8, 79740. [CrossRef]
4. Amarasinghe, A.; Kuritsky, J.N.; Letson, G.W.; Margolis, H.S. Dengue Virus Infection in Africa. Emerg. Infect. Dis. 2011, 17,
1349–1354. [CrossRef]
5. Krakower, D.S.; Mayer, K.H. Pre-Exposure Prophylaxis to Prevent HIV Infection: Current Status, Future Opportunities and
Challenges. Drugs 2015, 75, 243–251. [CrossRef]
Biosensors 2022, 12, 180 13 of 17

6. Lacroix, R.; Mukabana, W.R.; Gouagna, L.C.; Koella, J.C. Malaria Infection Increases Attractiveness of Humans to Mosquitoes.
PLoS Biol. 2005, 3, e298. [CrossRef]
7. Mori, R.; Lakhanpaul, M.; Verrier-Jones, K. Diagnosis and management of urinary tract infection in children: Summary of NICE
guidance. BMJ 2007, 335, 395–397. [CrossRef]
8. Saenz, R.A.; Quinlivan, M.; Elton, D.; MacRae, S.; Blunden, A.S.; Mumford, J.A.; Daly, J.M.; Digard, P.; Cullinane, A.; Grenfell,
B.T.; et al. Dynamics of Influenza Virus Infection and Pathology. J. Virol. 2010, 84, 3974–3983. [CrossRef]
9. Shereen, M.A.; Khan, S.; Kazmi, A.; Bashir, N.; Siddique, R. COVID-19 infection: Emergence, transmission, and characteristics of
human coronaviruses. J. Adv. Res. 2020, 24, 91–98. [CrossRef]
10. Amith, S.R.; Jayanth, P.; Franchuk, S.; Siddiqui, S.; Seyrantepe, V.; Gee, K.; Basta, S.; Beyaert, R.; Pshezhetsky, A.V.; Szewczuk,
M.R. Dependence of pathogen molecule-induced Toll-like receptor activation and cell function on Neu1 sialidase. Glycoconj. J.
2009, 26, 1197–1212. [CrossRef]
11. Bhatia, S.; Camacho, L.C.; Haag, R. Pathogen Inhibition by Multivalent Ligand Architectures. J. Am. Chem. Soc. 2016, 138,
8654–8666. [CrossRef]
12. Groom, J.; Richmond, J.; Murooka, T.; Sorensen, E.; Sung, J.H.; Bankert, K.; von Andrian, U.H.; Moon, J.J.; Mempel, T.R.; Luster,
A.D. CXCR3 Chemokine Receptor-Ligand Interactions in the Lymph Node Optimize CD4+ T Helper 1 Cell Differentiation.
Immunity 2012, 37, 1091–1103. [CrossRef]
13. Weekes, M.P.; Tomasec, P.; Huttlin, E.L.; Fielding, C.A.; Nusinow, D.; Stanton, R.J.; Wang, E.C.Y.; Aicheler, R.; Murrell, I.;
Wilkinson, G.W.G.; et al. Quantitative Temporal Viromics: An Approach to Investigate Host-Pathogen Interaction. Cell 2014, 157,
1460–1472. [CrossRef]
14. Lazcka, O.; Del Campo, F.J.; Munoz, F.X. Pathogen detection: A perspective of traditional methods and biosensors. Biosens.
Bioelectron. 2007, 22, 1205–1217. [CrossRef]
15. Kang, J.; Kim, M.G. Advancements in DNA-assisted Immunosensors. Biochip J. 2020, 14, 18–31. [CrossRef]
16. Verma, J.; Saxena, S.; Babu, S.G. ELISA-Based Identification and Detection of Microbes, in Analyzing Microbes: Manual of Molecular
Biology Techniques; Arora, D.K., Das, S., Sukumar, M., Eds.; Springer: Berlin/Heidelberg, Germany, 2013; pp. 169–186.
17. Cui, N.; Su, S.; Sun, P.; Zhang, Y.; Han, N.; Cui, Z. Isolation and pathogenic analysis of virulent Marek’s disease virus field strain
in China. Poult. Sci. 2016, 95, 1521–1528. [CrossRef]
18. Eparvier, A.; Alabouvette, C. Use of ELISA and GUS-transformed strains to study competition between pathogenic and non-
pathogenic Fusarium oxysporum for root colonization. Biocontrol. Sci. Technol. 1994, 4, 35–47. [CrossRef]
19. Ho, Y.-P.; Reddy, P.M. Identification of Pathogens by Mass Spectrometry. Clin. Chem. 2010, 56, 525–536. [CrossRef]
20. K’Owino, I.O.; Sadik, O.A. Impedance Spectroscopy: A Powerful Tool for Rapid Biomolecular Screening and Cell Culture
Monitoring. Electroanalysis 2005, 17, 2101–2113. [CrossRef]
21. Li, Y.; Cu, Y.T.H.; Luo, D. Multiplexed detection of pathogen DNA with DNA-based fluorescence nanobarcodes. Nat. Biotechnol.
2005, 23, 885–889. [CrossRef]
22. Louws, F.; Rademaker, J.; de Bruijn, F. The Three ds of PCR-based genomic analysis of phytobacteria: Diversity, Detection, and
Disease Diagnosis. Annu. Rev. Phytopathol. 1999, 37, 81–125. [CrossRef] [PubMed]
23. Patel, J.B. 16S rRNA Gene Sequencing for Bacterial Pathogen Identification in the Clinical Laboratory. J. Mol. Diagn. 2001, 6,
313–321. [CrossRef]
24. Pruzzo, C.; Guzmán, C.A.; Dainelli, B. Incidence of hemagglutination activity among pathogenic and non-pathogenic Bacteroides
fragilis strains and role of capsule and pili in HA and adherence. FEMS Microbiol. Lett. 1989, 59, 113–118. [CrossRef]
25. Tylewska-Wierzbanowska, S.; Chmielewski, T. Limitation of serological testing for Lyme borreliosis: Evaluation of ELISA and
western blot in comparison with PCR and culture methods. Wien. Klin. Wochenschr. 2002, 114, 601–605. [PubMed]
26. Morita, Y.; Sakaguchi, T.; Unno, N.; Shibasaki, Y.; Suzuki, A.; Fukumoto, K.; Inaba, K.; Baba, S.; Takehara, Y.; Suzuki, S.; et al.
Detection of hepatocellular carcinomas with near-infrared fluorescence imaging using indocyanine green: Its usefulness and
limitation. Int. J. Clin. Oncol. 2011, 18, 232–241. [CrossRef]
27. Mayfield, S.; Lopata, A.L.; Branch, G.M. Limitation and failure of immunological technique (ELISA) in resolving the diet of the
South African rock lobster Jasus Ialandii. Mar. Biol. 2000, 137, 595–604. [CrossRef]
28. Diamandis, E.P. Mass spectrometry as a diagnostic and a cancer biomarker discovery tool: Opportunities and potential limitations.
Mol. Cell. Proteom. 2004, 3, 367–378. [CrossRef]
29. Bélec, L.; Authier, J.R.; Eliezer-Vanerot, M.C.; Piédouillet, C.; Mohamed, A.S.; Gherardi, R.K. Myoglobin as a polymerase
chain reaction (PCR) inhibitor: A limitation for PCR from skeletal muscle tissue avoided by the use of Thermus thermophilus
polymerase. Muscle Nerve 1998, 21, 1064–1067. [CrossRef]
30. Abicht, A.; Dusl, M.; Gallenmüller, C.; Guergueltcheva, V.; Schara, U.; Della Marina, A.; Wibbeler, E.; Almaras, S.; Mihaylova, V.;
von der Hagen, M.; et al. Congenital myasthenic syndromes: Achievements and limitations of phenotype-guided gene-after-gene
sequencing in diagnostic practice: A study of 680 patients. Hum. Mutat. 2012, 33, 1474–1484. [CrossRef]
31. Singh, P.; Onodera, T.; Mizuta, Y.; Matsumoto, K.; Miura, N.; Toko, K. Dendrimer modified biochip for detection of 2,4,6
trinitrotoluene on SPR immunosensor: Fabrication and advantages. Sens. Actuators B Chem. 2009, 137, 403–409. [CrossRef]
32. Brolo, A.G. Plasmonics for future biosensors. Nat. Photon. 2012, 6, 709–713. [CrossRef]
33. Shrivastav, A.M.; Cvelbar, U.; Abdulhalim, I. A comprehensive review on plasmonic-based biosensors used in viral diagnostics.
Commun. Biol. 2021, 4, 70. [CrossRef]
Biosensors 2022, 12, 180 14 of 17

34. Chain, C.Y.; Millone, M.A.D.; Cisneros, J.S.; Ramirez, E.A.; Vela, M.E. Surface Plasmon Resonance as a Characterization Tool for
Lipid Nanoparticles Used in Drug Delivery. Front. Chem. 2021, 8, 605307. [CrossRef]
35. Nguyen, H.H.; Park, J.; Kang, S.; Kim, M. Surface Plasmon Resonance: A Versatile Technique for Biosensor Applications. Sensors
2015, 15, 10481–10510. [CrossRef]
36. Huang, H.-T.; Huang, C.Y.; Ger, T.-R.; Wei, Z.-H. Anti-integrin and integrin detection using the heat dissipation of surface
plasmon resonance. Appl. Phys. Lett. 2013, 102, 111109. [CrossRef]
37. Kim, J.; Chun, S.H.; Amornkitbamrung, L.; Song, C.; Yuk, J.S.; Ahn, S.Y.; Kim, B.W.; Lim, Y.T.; Oh, B.K.; Um, S.H. Gold nanoparticle
clusters for the investigation of therapeutic efficiency against prostate cancer under near-infrared irradiation. Nano Converg. 2020,
7, 5. [CrossRef]
38. Kalogianni, D.P. Nanotechnology in emerging liquid biopsy applications. Nano Converg. 2021, 8, 13. [CrossRef]
39. Chen, Y.; Ai, B.; Wong, Z.J. Soft optical metamaterials. Nano Converg. 2020, 7, 18. [CrossRef]
40. Prodanov, M.F.; Pogorelova, N.V.; Kryshtal, A.P.; Klymchenko, A.S.; Mely, Y.; Semynozhenko, V.P.; Krivoshey, A.I.; Reznikov, Y.A.;
Yarmolenko, S.N.; Goodby, J.W.; et al. Thermodynamically Stable Dispersions of Quantum Dots in a Nematic Liquid Crystal.
Langmuir 2013, 29, 9301–9309. [CrossRef]
41. Wadhwa, S.; John, A.T.; Nagabooshanam, S.; Mathur, A.; Narang, J. Graphene quantum dot-gold hybrid nanoparticles inte-
grated aptasensor for ultra-sensitive detection of vitamin D3 towards point-of-care application. Appl. Surf. Sci. 2020, 521,
146427. [CrossRef]
42. Dahlin, A.B. Sensing applications based on plasmonic nanopores: The hole story. Analyst 2015, 140, 4748–4759. [CrossRef] [PubMed]
43. Gupta, B.D.; Verma, R.K. Surface Plasmon Resonance-Based Fiber Optic Sensors: Principle, Probe Designs, and Some Applications.
J. Sens. 2009, 2009, 979761. [CrossRef]
44. Shinn, M.; Robertson, W. Surface plasmon-like sensor based on surface electromagnetic waves in a photonic band-gap material.
Sens. Actuators B Chem. 2005, 105, 360–364. [CrossRef]
45. Coskun, A.F.; Cetin, A.E.; Galarreta, B.; Alvarez, D.A.; Altug, H.; Ozcan, A. Lensfree optofluidic plasmonic sensor for real-time
and label-free monitoring of molecular binding events over a wide field-of-view. Sci. Rep. 2014, 4, 6789. [CrossRef] [PubMed]
46. Erdem, Ö.; Saylan, Y.; Cihangir, N.; Denizli, A. Molecularly imprinted nanoparticles based plasmonic sensors for real-time
Enterococcus faecalis detection. Biosens. Bioelectron. 2018, 126, 608–614. [CrossRef]
47. Soelberg, S.D.; Chinowsky, T.; Geiss, G.; Spinelli, C.B.; Stevens, R.; Near, S.; Kauffman, P.; Yee, S.; Furlong, C.E. A portable
surface plasmon resonance sensor system for real-time monitoring of small to large analytes. J. Ind. Microbiol. Biotechnol. 2005, 32,
669–674. [CrossRef]
48. Kannan, R.; Rahing, V.; Cutler, C.; Pandrapragada, R.; Katti, K.K.; Kattumuri, V.; Robertson, J.D.; Casteel, S.J.; Jurisson, S.; Smith,
C.; et al. Nanocompatible Chemistry toward Fabrication of Target-Specific Gold Nanoparticles. J. Am. Chem. Soc. 2006, 128,
11342–11343. [CrossRef]
49. Kawawaki, T.; Takahashi, Y.; Tatsuma, T. Enhancement of Dye-Sensitized Photocurrents by Gold Nanoparticles: Effects of
Plasmon Coupling. J. Phys. Chem. C 2013, 117, 5901–5907. [CrossRef]
50. Nooke, A.; Beck, U.; Hertwig, A.; Krause, A.; Krüger, H.; Lohse, V.; Negendank, D.; Steinbach, J. On the application of gold based
SPR sensors for the detection of hazardous gases. Sens. Actuators B Chem. 2010, 149, 194–198. [CrossRef]
51. Patnaik, A.; Senthilnathan, K.; Jha, R. Graphene-Based Conducting Metal Oxide Coated D-Shaped Optical Fiber SPR Sensor. IEEE
Photon. Technol. Lett. 2015, 27, 2437–2440. [CrossRef]
52. Patra, C.R.; Bhattacharya, R.; Mukhopadhyay, D.; Mukherjee, P. Fabrication of gold nanoparticles for targeted therapy in
pancreatic cancer. Adv. Drug Deliv. Rev. 2010, 62, 346–361. [CrossRef]
53. Sexton, B.; Feltis, B.; Davis, T. Characterisation of gold surface plasmon resonance sensor substrates. Sens. Actuators A Phys. 2008,
141, 471–475. [CrossRef]
54. Sugunan, A.; Thanachayanont, C.; Dutta, J.; Hilborn, J. Heavy-metal ion sensors using chitosan-capped gold nanoparticles. Sci.
Technol. Adv. Mater. 2005, 6, 335–340. [CrossRef]
55. Zhang, Y.; Liang, P.; Wang, Y.; Zhang, Y.; Liu, Z.; Wei, Y.; Zhu, Z.; Zhao, E.; Yang, J.; Yuan, L. Cascaded distributed multichannel
fiber SPR sensor based on gold film thickness adjustment approach. Sens. Actuator A Phys. 2017, 267, 526–531. [CrossRef]
56. Jung, J.H.; Cheon, D.S.; Liu, F.; Lee, K.B.; Seo, T.S. A Graphene Oxide Based Immuno-biosensor for Pathogen Detection. Angew.
Chem. Int. Ed. 2010, 49, 5708–5711. [CrossRef]
57. Liu, X.; Shi, L.; Jiang, W.; Zhang, J.; Huang, L. Taking full advantage of KMnO4 in simplified Hummers method: A green and one
pot process for the fabrication of alpha MnO2 nanorods on graphene oxide. Chem. Eng. Sci. 2018, 192, 414–421. [CrossRef]
58. Liu, Y.; Yu, D.; Zeng, C.; Miao, Z.; Dai, L. Biocompatible Graphene Oxide-Based Glucose Biosensors. Langmuir 2010, 26,
6158–6160. [CrossRef]
59. Szunerits, S.; Maalouli, N.; Wijaya, E.; Vilcot, J.-P.; Boukherroub, R. Recent advances in the development of graphene-based
surface plasmon resonance (SPR) interfaces. Anal. Bioanal. Chem. 2013, 405, 1435–1443. [CrossRef]
60. Cai, B.; Huang, L.; Zhang, H.; Sun, Z.; Zhang, Z.; Zhang, G.-J. Gold nanoparticles-decorated graphene field-effect transistor
biosensor for femtomolar MicroRNA detection. Biosens. Bioelectron. 2015, 74, 329–334. [CrossRef]
61. Song, Y.; Xu, T.; Xu, L.-P.; Zhang, X. Nanodendritic gold/graphene-based biosensor for tri-mode miRNA sensing. Chem. Commun.
2019, 55, 1742–1745. [CrossRef]
Biosensors 2022, 12, 180 15 of 17

62. Zhang, L.; Han, G.; Liu, Y.; Tang, J.; Tang, W. Immobilizing haemoglobin on gold/graphene–chitosan nanocomposite as efficient
hydrogen peroxide biosensor. Sens. Actuators B Chem. 2014, 197, 164–171. [CrossRef]
63. Ghosh, S.K.; Pal, T. Interparticle Coupling Effect on the Surface Plasmon Resonance of Gold Nanoparticles: From Theory to
Applications. Chem. Rev. 2007, 107, 4797–4862. [CrossRef] [PubMed]
64. Su, Q.; Xue, T.; Zhang, Y.; Lan, K.; Zou, Q. Fabrication of enhanced silver nanowire films via self-assembled gold nanoparticles
without post-treatment. Mater. Lett. 2018, 236, 218–221. [CrossRef]
65. Bianco, M.; Sonato, A.; De Girolamo, A.; Pascale, M.; Romanato, F.; Rinaldi, R.; Arima, V. An aptamer-based SPR-polarization
platform for high sensitive OTA detection. Sens. Actuators B Chem. 2017, 241, 314–320. [CrossRef]
66. Luo, Z.; Zhang, J.; Wang, Y.; Chen, J.; Li, Y.; Duan, Y. An aptamer based method for small molecules detection through
monitoring salt-induced AuNPs aggregation and surface plasmon resonance (SPR) detection. Sens. Actuators B Chem. 2016, 236,
474–479. [CrossRef]
67. Sun, L.; Wu, L.; Zhao, Q. Aptamer based surface plasmon resonance sensor for aflatoxin B1. Mikrochim. Acta 2017, 184,
2605–2610. [CrossRef]
68. Wang, W.; Chen, C.; Qian, M.; Zhao, X.S. Aptamer biosensor for protein detection using gold nanoparticles. Anal. Biochem. 2008,
373, 213–219. [CrossRef]
69. Liu, C.; Wang, J.; Wang, F.; Su, W.; Yang, L.; Lv, J.; Fu, G.; Li, X.; Liu, Q.; Sun, T.; et al. Surface plasmon resonance (SPR) infrared
sensor based on D-shape photonic crystal fibers with ITO coatings. Opt. Commun. 2020, 464, 125496. [CrossRef]
70. Shibayama, J. Three-Dimensional Numerical Investigation of an Improved Surface Plasmon Resonance Waveguide Sensor. IEEE
Photon. Technol. Lett. 2010, 22, 643–645. [CrossRef]
71. Wang, W.; Cui, H. Chitosan-Luminol Reduced Gold Nanoflowers: From One-Pot Synthesis to Morphology-Dependent SPR and
Chemiluminescence Sensing. J. Phys. Chem. C 2008, 112, 10759–10766. [CrossRef]
72. Karthik, V.; Selvakumar, P.; Kumar, P.S.; Vo, D.-V.N.; Gokulakrishnan, M.; Keerthana, P.; Elakkiya, V.T.; Rajeswari, R. Graphene-
based materials for environmental applications: A review. Environ. Chem. Lett. 2021, 19, 3631–3644. [CrossRef]
73. Yan, Z.; Xue, H.; Berning, K.; Lam, Y.-W.; Lee, C.-S. Identification of Multifunctional Graphene–Gold Nanocomposite for
Environment-Friendly Enriching, Separating, and Detecting Hg2+ Simultaneously. ACS Appl. Mater. Interfaces 2014, 6, 22761–
22768. [CrossRef]
74. Kodoyianni, V. Label-free analysis of biomolecular interactions using SPR imaging. Biotechniques 2011, 50, 32–40. [CrossRef]
75. Slavık, R.; Homola, J.; Brynda, E. A miniature fiber optic surface plasmon resonance sensor for fast detection of staphylococcal
enterotoxin B. Biosens. Bioelectron. 2002, 17, 591–595. [CrossRef]
76. Raikwar, S.; Prajapati, Y.K.; Srivastava, D.K.; Maurya, J.B.; Saini, J.P. Detection of Leptospirosis Bacteria in Rodent Urine by
Surface Plasmon Resonance Sensor Using Graphene. Photon. Sens. 2020, 11, 305–313. [CrossRef]
77. Mudgal, N.; Yupapin, P.; Ali, J.; Singh, G. BaTiO3 -Graphene-Affinity Layer–Based Surface Plasmon Resonance (SPR) Biosensor
for Pseudomonas Bacterial Detection. Plasmonics 2020, 15, 1221–1229. [CrossRef]
78. Omar, N.A.S.; Fen, Y.W.; Abdullah, J.; Zaid, M.H.M.; Daniyal, W.M.E.M.M.; Mahdi, M.A. Sensitive surface plasmon resonance
performance of cadmium sulfide quantum dots-amine functionalized graphene oxide based thin film towards dengue virus
E-protein. Opt. Laser Technol. 2019, 114, 204–208. [CrossRef]
79. Bong, J.H.; Kim, T.H.; Jung, J.; Lee, S.J.; Sung, J.S.; Lee, C.K.; Kang, M.-J.; Kim, H.O.; Pyun, J.C. Pig Sera-derived Anti-SARS-CoV-2
Antibodies in Surface Plasmon Resonance Biosensors. Biochip J. 2020, 14, 358–368. [CrossRef]
80. Masdor, N.A.; Altintas, Z.; Tothill, I.E. Surface Plasmon Resonance Immunosensor for the Detection of Campylobacter jejuni.
Chemosenso 2017, 5, 16. [CrossRef]
81. Funari, R.; Chu, K.Y.; Shen, A.Q. Detection of antibodies against SARS-CoV-2 spike protein by gold nanospikes in an opto-
microfluidic chip. Biosens. Bioelectron 2020, 169, 112578. [CrossRef]
82. Vázquez-Guardado, A.; Mehta, F.; Jimenez, B.; Biswas, A.; Ray, K.; Baksh, A.; Lee, S.; Saraf, N.; Seal, S.; Chanda, D. DNA-
Modified Plasmonic Sensor for the Direct Detection of Virus Biomarkers from the Blood. Nano Lett. 2021, 21, 7505–7511.
[CrossRef] [PubMed]
83. Omar, N.A.S.; Fen, Y.W.; Saleviter, S.; Daniyal, W.M.E.M.M.; Anas, N.A.A.; Ramdzan, N.S.M.; Roshidi, M.D.A. Development of a
Graphene-Based Surface Plasmon Resonance Optical Sensor Chip for Potential Biomedical Application. Materials 2019, 12, 1928.
[CrossRef] [PubMed]
84. Kim, J.-W.; Kim, M.; Lee, K.K.; Chung, K.H.; Lee, C.-S. Effects of Graphene Oxide-Gold Nanoparticles Nanocomposite on Highly
Sensitive Foot-and-Mouth Disease Virus Detection. Nanomaterials 2020, 10, 1921. [CrossRef] [PubMed]
85. Liu, M.; Zheng, C.; Cui, M.; Zhang, X.; Yang, D.-P.; Wang, X.; Cui, D. Graphene oxide wrapped with gold nanorods as a tag in a
SERS based immunoassay for the hepatitis B surface antigen. Mikrochim. Acta 2018, 185, 458. [CrossRef]
86. Song, M.-S.; Sekhon, S.S.; Shin, W.-R.; Rhee, S.-K.; Ko, J.H.; Kim, S.Y.; Min, J.; Ahn, J.-Y.; Kim, Y.-H. Aptamer-Immobilized Surface
Plasmon Resonance Biosensor for Rapid and Sensitive Determination of Virulence Determinant. J. Nanosci. Nanotechnol. 2018, 18,
3095–3101. [CrossRef]
87. Shin, W.-R.; Sekhon, S.S.; Rhee, S.-K.; Ko, J.H.; Ahn, J.-Y.; Min, J.; Kim, Y.-H. Aptamer-Based Paper Strip Sensor for Detecting
Vibrio fischeri. ACS Comb. Sci. 2018, 20, 261–268. [CrossRef]
88. Kim, S.; Lee, S.; Lee, H.J. An aptamer-aptamer sandwich assay with nanorod-enhanced surface plasmon resonance for attomolar
concentration of norovirus capsid protein. Sens. Actuators B Chem. 2018, 273, 1029–1036. [CrossRef]
Biosensors 2022, 12, 180 16 of 17

89. Chuang, C.-S.; Wu, C.-Y.; Juan, P.-H.; Hou, N.-C.; Fan, Y.-J.; Wei, P.-K.; Sheen, H.-J. LMP1 gene detection using a capped gold
nanowire array surface plasmon resonance sensor in a microfluidic chip. Analyst 2020, 145, 52–60. [CrossRef]
90. Rippa, M.; Castagna, R.; Brandi, S.; Fusco, G.; Monini, M.; Chen, D.; Zhou, J.; Zyss, J.; Petti, L. Octupolar Plasmonic Nanosensor
Based on Ordered Arrays of Triangular Au Nanopillars for Selective Rotavirus Detection. ACS Appl. Nano Mater. 2020, 3,
4837–4844. [CrossRef]
91. Kaushik, S.; Tiwari, U.K.; Pal, S.S.; Sinha, R.K. Rapid detection of Escherichia coli using fiber optic surface plasmon reso-
nance immunosensor based on biofunctionalized Molybdenum disulfide (MoS2 ) nanosheets. Biosens. Bioelectron. 2019, 126,
501–509. [CrossRef]
92. Das, S.; Singh, S.; Singh, V.; Joung, D.; Dowding, J.M.; Reid, D.; Anderson, J.; Zhai, L.; Khondaker, S.I.; Self, W.T.; et al. Oxygenated
Functional Group Density on Graphene Oxide: Its Effect on Cell Toxicity. Part. Part. Syst. Charact. 2013, 30, 148–157. [CrossRef]
93. Gupta, B.; Kumar, N.; Panda, K.; Kanan, V.; Joshi, S.; Visoly-Fisher, I. Role of oxygen functional groups in reduced graphene oxide
for lubrication. Sci. Rep. 2017, 7, srep45030. [CrossRef]
94. Zhao, J.; Wang, Y.; Liao, C.; Cao, S.; Li, M.; Wang, Y. Graphene oxide modified surface plasmon resonance sensor based on
side-polished fiber. In Proceedings of the 2017 25th Optical Fiber Sensors Conference (OFS), Jeju, Korea, 24–28 April 2017.
95. Xia, Z.; Chu, F.; Bian, Z.; Zhang, Z.; Li, J.; Guo, Z. Study of Surface Plasmon Resonance Sensor Based on Polymer-Tipped Optical
Fiber With Barium Titanate Layer. J. Light. Technol. 2020, 38, 912–918. [CrossRef]
96. Anas, N.A.A.; Fen, Y.W.; Yusof, N.A.; Omar, N.A.S.; Daniyal, W.M.E.M.M.; Ramdzan, N.S.M. Highly sensitive surface plasmon
resonance optical detection of ferric ion using CTAB/hydroxylated graphene quantum dots thin film. J. Appl. Phys. 2020, 128,
83105. [CrossRef]
97. Zhang, H.; Song, D.; Gao, S.; Zhang, J.; Zhang, H.; Sun, Y. Novel SPR biosensors based on metal nanoparticles decorated with
graphene for immunoassay. Sens. Actuators B Chem. 2013, 188, 548–554. [CrossRef]
98. Bodrumlu, E. Biocompatibility of retrograde root filling materials: A review. Aust. Endod. J. 2008, 34, 30–35. [CrossRef]
99. Haume, K.; Rosa, S.; Grellet, S.; Śmiałek, M.A.; Butterworth, K.T.; Solov’Yov, A.V.; Prise, K.M.; Golding, J.; Mason, N.J. Gold
nanoparticles for cancer radiotherapy: A review. Cancer Nanotechnol. 2016, 7, 8. [CrossRef]
100. Panikkanvalappil, S.R.; Hooshmand, N.; El-Sayed, M.A. Intracellular Assembly of Nuclear-Targeted Gold Nanosphere Enables
Selective Plasmonic Photothermal Therapy of Cancer by Shifting Their Absorption Wavelength toward Near-Infrared Region.
Bioconjugate Chem. 2017, 28, 2452–2460. [CrossRef]
101. Gramotnev, D.K.; Vogel, M.W. Ultimate capabilities of sharp metal tips for plasmon nanofocusing, near-field trapping and sensing.
Phys. Lett. A 2011, 375, 3464–3468. [CrossRef]
102. Yoo, D.; Barik, A.; de León-Pérez, F.; Mohr, D.A.; Pelton, M.; Martín-Moreno, L.; Oh, S.-H. Plasmonic Split-Trench Resonator for
Trapping and Sensing. ACS Nano 2021, 15, 6669–6677. [CrossRef]
103. Zaman, M.A.; Padhy, P.; Hesselink, L. Near-field optical trapping in a non-conservative force field. Sci. Rep. 2019, 9, 649. [CrossRef]
104. Padhy, P.; Zaman, M.A.; Hansen, P.; Hesselink, L. On the substrate contribution to the back action trapping of plasmonic
nanoparticles on resonant near-field traps in plasmonic films. Opt. Express 2017, 25, 26198–26214. [CrossRef]
105. Wang, K.; Schonbrun, E.; Steinvurzel, P.; Crozier, K. Trapping and rotating nanoparticles using a plasmonic nano-tweezer with an
integrated heat sink. Nat. Commun. 2011, 2, 469. [CrossRef]
106. Zaman, M.A.; Padhy, P.; Hesselink, L. Solenoidal optical forces from a plasmonic Archimedean spiral. Phys. Rev. A 2019, 100,
13857. [CrossRef] [PubMed]
107. Islam, S.; Islam, M.R.; Sultana, J.; Dinovitser, A.; Ng, B.W.-H.; Abbott, D. Exposed-core localized surface plasmon resonance
biosensor. J. Opt. Soc. Am. B 2019, 36, 2306–2311. [CrossRef]
108. Wu, L.; Chu, H.S.; Koh, W.S.; Li, E.P. Highly sensitive graphene biosensors based on surface plasmon resonance. Opt. Express
2010, 18, 14395–14400. [CrossRef] [PubMed]
109. Anas, N.A.A.; Fen, Y.W.; Omar, N.A.S.; Daniyal, W.M.E.M.M.; Ramdzan, N.S.M.; Saleviter, S. Development of Graphene Quantum
Dots-Based Optical Sensor for Toxic Metal Ion Detection. Sensors 2019, 19, 3850. [CrossRef] [PubMed]
110. Nayak, J.K.; Parhi, P.; Jha, R. Graphene oxide encapsulated gold nanoparticle based stable fibre optic sucrose sensor. Sens.
Actuators B Chem. 2015, 221, 835–841. [CrossRef]
111. Kim, S.M.; Kim, J.; Noh, S.; Sohn, H.; Lee, T. Recent Development of Aptasensor for Influenza Virus Detection. Biochip J. 2020, 14,
327–339. [CrossRef]
112. Lee, G.-H.; Hah, S.S. Coomassie blue is sufficient for specific protein detection of aptamer-conjugated chips. Bioorganic Med. Chem.
Lett. 2012, 22, 1520–1522. [CrossRef]
113. Stadtherr, K.; Wolf, A.H.; Lindner, P. An Aptamer-Based Protein Biochip. Anal. Chem. 2005, 77, 3437–3443. [CrossRef]
114. Smirnov, I.; Shafer, R.H. Effect of Loop Sequence and Size on DNA Aptamer Stability. Biochemistry 2000, 39, 1462–1468. [CrossRef]
115. Hasegawa, H.; Savory, N.; Abe, K.; Ikebukuro, K. Methods for Improving Aptamer Binding Affinity. Molecules 2016, 21,
421. [CrossRef]
116. Miller, O.D.; Ilic, O.; Christensen, T.; Reid, M.T.H.; Atwater, H.A.; Joannopoulos, J.D.; Soljačić, M.; Johnson, S.G. Limits to the
Optical Response of Graphene and Two-Dimensional Materials. Nano Lett. 2017, 17, 5408–5415. [CrossRef]
117. Kant, R.; Tabassum, R.; Gupta, B.D. Integrating nanohybrid membranes of reduced graphene oxide: Chitosan: Silica sol gel with
fiber optic SPR for caffeine detection. Nanotechnology 2017, 28, 195502. [CrossRef]
Biosensors 2022, 12, 180 17 of 17

118. Singh, M.; Holzinger, M.; Tabrizian, M.; Winters, S.; Berner, N.C.; Cosnier, S.; Duesberg, G.S. Noncovalently Functionalized
Monolayer Graphene for Sensitivity Enhancement of Surface Plasmon Resonance Immunosensors. J. Am. Chem. Soc. 2015, 137,
2800–2803. [CrossRef]
119. Verma, R.; Gupta, B.D.; Jha, R. Sensitivity enhancement of a surface plasmon resonance based biomolecules sensor using graphene
and silicon layers. Sens. Actuators B Chem. 2011, 160, 623–631. [CrossRef]
120. Mishra, A.K.; Mishra, S.K.; Verma, R.K. Graphene and Beyond Graphene MoS2: A New Window in Surface-Plasmon-Resonance-
Based Fiber Optic Sensing. J. Phys. Chem. C 2016, 120, 2893–2900. [CrossRef]