Development of Integrated Electrochemical–Quartz Crystal

Microbalance Biosensor Arrays: Towards Ultrasensitive, Multiplexed

and Rapid Point-of-Care Dengue Detection

Ahmad Anwar Zainuddin

, Anis Nurashikin Nordin

, Mohd Afiq Mohd Asri

Rosminazuin Ab Rahim

, Cyril Guines

, Matthieu Chatras

Arnaud Pothier

and Wing Cheung Mak

Department of Electrical & Computer Engineering, International Islamic University Malaysia, Kuala Lumpur, Malaysia

XLIM Research Institute, UMR CNRS 7252 Université de Limoges, Limoges Cedex, France

Biosensors and Bioelectronics Centre, Department of Physics, Chemistry and Biology (IFM),

Linkoping University, 58183, Linkoping, Sweden

Keywords: Dengue, Biosensor, Sensor Arrays, Integrated Electrochemical-Quartz Crystal Microbalance, Point-of-Care

Diagnostics.

Abstract: Dengue is an infectious mosquito-borne viral disease that affects approximately 50 million people annually

worldwide and is prevalent mostly in the tropics. Severe cases of dengue can be fatal, making early detection

and fast diagnosis crucial towards improving patient care and survival rates. Currently, early detection can be

achieved through detection of NS1 protein, using ELISA technique. Unfortunately, ELISA is an expensive

method, making it unsuitable as a screening technique, especially in low-resource settings. In this work, we

present a prototype device and its early validation studies, of an integrated electrochemical and mass-sensor

for dengue NS1 antigen. The sensor is connected to open source mass-sensing software and hardware,

OpenQCM which makes it easily portable. Having dual-measurement capabilities (mass and impedance)

increases the sensitivity of the sensor. Preliminary studies suggest that the prototype could achieve ultralow

limit of detection as low as 10 ng mL

-1

, dual-sensing cross-validation capability, portable size, sample-to-

analysis time of less than 30 minutes, and parallelization of multiple assays. This work could lead to early

and accurate dengue detection in routine point-of-care settings.

1 INTRODUCTION

Dengue is an infectious tropical disease transmitted

by Aedes mosquitoes, which pose serious health

threats, especially in tropical and subtropical regions.

Dengue virus has four serotypes (DENV 1–4), which

produces different symptoms starting from mild fever

to the more severe forms, which may lead to fatal

illnesses such as dengue hemorrhagic fever (DHF)

and dengue shock syndrome (DSS). Malaysia is

among the seriously affected countries with a rapid

increase in the number of dengue fever cases from

43,346 with 92 deaths in 2013, to 101,357 cases

including 237 deaths in 2016 (Ahmad et al., 2018;

Shafie et al., 2017). In Indonesia, there was a reported

average of 94 564 cases and between 472 and 1446

deaths per year in between year of 2001 and 2011

(Wahyono et al., 2017). On the other hand, three

serotypes of dengue virus (1, 2 and 3) detection are

circulating in Saudi Arabia where 1790 confirmed

dengue cases between 2005 and 2016 with highest

outbreaks in 2016 (555 cases) (Mohammed et al.,

2018). In Africa, outbreaks of DF are increasing in

size and frequency but are not being consistently

reported to the WHO.

NS1 (non-structural) dengue protein antigen

capture immunoassays, have been widely used as a

dengue fever biomarker for the early stage detection

(up to 7 days) in dengue diagnosis (Cecchetto et al.,

2015). The NS1 protein is conserved among all four

dengue serotypes, and is expressed or secreted by

infected host cells (Masrinoul et al., 2011). A

practical diagnostic titre in serum sample of human

bodies can range from 40 ng mL

-1

to 2000 ng mL

-1

(Dias et al., 2013). The combination of the NS1

antigen and specific antibody tests such as IgM and

IgG are popularly discussed to enhance the diagnostic

efficiency for early diagnosis of dengue infection

220

Zainuddin, A., Nordin, A., Asri, M., Rahim, R., Guines, C., Chatras, M., Pothier, A. and Mak, W.

Development of Integrated Electrochemical–Quartz Crystal Microbalance Biosensor Arrays: Towards Ultrasensitive, Multiplexed and Rapid Point-of-Care Dengue Detection.

DOI: 10.5220/0007523802200227

In Proceedings of the 12th International Joint Conference on Biomedical Engineering Systems and Technologies (BIOSTEC 2019), pages 220-227

ISBN: 978-989-758-353-7

(Casenghi et al., 2014; Kassim et al., 2011; Omar and

Fen, 2018). It was also previously shown that NS1

also presents itself in asymptomatic patients (Ashshi,

2017).

Recently, lab-on-chip (LoC) devices have been

increasingly exploited for biosensor applications due

to its ability to execute comprehensive laboratory

protocols on a single integrated miniature MEMS

device, with lower time and reagent consumption

than a normal laboratory protocol. Biosensing

techniques such as electrochemistry (Electrochemical

Impedance Spectroscopy, EIS) and mass sensing

(Quartz Crystal Microbalance, QCM) measurement

have been extensively used due to their high

sensitivity and are easy to miniaturize for use in point-

of-care (PoC) applications (Pal et al., 2015). Among

emerging applications of PoC biosensors include

dengue virus detection for clinical screening and

diagnosis. Many of these biosensors rely on

immunoaffinity principles, similar to conventional

enzyme-linked immunosorbent assay (ELISA),

where antigen-specific antibody is immobilized on a

solid sensor surface. In typical clinical practice,

ELISA is used for dengue diagnosis; complemented

by virus isolation, RT-PCR and serology methods.

These methods however are time-consuming, and

require tedious steps, expensive instrumentations and

trained personnel (Darwish et al., 2015).

QCM measures minuscule changes in mass on the

surface of its sensor through the shifts in oscillation

and resonant frequency due to the damping effects

that occurs when analytes are deposited onto sensor

surface, as alternating current (AC) is subjected

through the piezoelectric quartz substrate (Liu et al.,

2013a). QCM is favoured in applications requiring

high sensitivities to microscale perturbations. The

typical configuration of QCMs is as single element

sensors, but a multichannel QCM array (MQCMA)

on the same crystal has been recently proposed (Liu

et al., 2013a; Zhang et al., 2017).

Electrochemical biosensors measure

perturbations in current or impedance on electrode

surface due to biological redox reaction or surface

interactions (Wang, 2006). One of the earliest

commercial examples of electrochemical biosensor is

the glucometer, which was introduced by Clark et al.

in 1962, in which screen-printed electrodes are

modified with glucose oxidase enzyme as a bio-

receptor (Silva et al., 2017). Electrochemical

biosensors are often preferred in development of

point-of-care applications due to its relatively low

cost, high sensitivity, and multiplexing capabilities

(Cecchetto et al., 2015).

Integrated sensors employing QCM and

electrochemical measurements are known as

electrochemical quartz crystal microbalance sensors

(EQCMs). These sensors are the combination of

detection methods which includes simultaneous

monitoring of mass change performed in parallel with

cyclic voltammetry (CV) and/or electrochemical

impedance spectroscopy (EIS) measurements on

common electrodes. In EQCM, the electrodes serve

both as QCM excitation electrodes, as well as the

electrochemical working electrode. The counter and

reference electrodes are added on one side of the

quartz surface (Xiao and Zeng, 2013). Several works

have been demonstrated for EQCM biosensors,

highlighting capabilities for higher sensitivity from

the QCM, higher selectivity and linear range from

electrochemistry, and cross-validation for accuracy

(Ma et al., 2015). More development is required

towards practical applications of EQCM in point-of-

care settings, however it offers a promising avenue

for new clinical insights.

In this position paper, we present a preliminary

work towards an ultrasensitive and rapid point-of-

care (PoC) device for early detection of dengue, based

on integrated EQCM. In section 2, the working

principles that form the basis of the EQCM sensor are

elaborated. We present in section 3 the development

of a multichannel EQCM array that serve as an early

functional prototype for a future integrated EQCM

(IEQCM) PoC device. We also show that it can

perform basic biosensing tests for dengue virus

antigens. In section 4, we analyze the potential

scientific and socio-economic impact of this

innovation, should it be successfully realized and

brought into clinical practices.

2 PRINCIPLES

2.1 Device Operation

To determine the optimal design of the integrated

biosensor, the principle of Quartz Crystal

Microbalance (QCM) and the electrochemistry

system is used here. Figure 1 depicts the geometrical

design and cross section of integrated biosensor.

The AC signal is applied to the top-bottom

electrode of AT-quartz to generate acoustic wave

energy (resonance frequency). Concurrently, for

electrochemistry detection typically consisting of

three metal electrodes: working, counter and

reference (Regiart et al., 2016). AC/DC signal is

excited between the two working and counter

electrodes; in which, the electric field will be induced

Development of Integrated Electrochemical–Quartz Crystal Microbalance Biosensor Arrays: Towards Ultrasensitive, Multiplexed and Rapid

Point-of-Care Dengue Detection

221

on these electrodes by the electric potential when a

probe marker or electrolyte is introduced into the

system (Mansor and Ibrahim, 2016). Thus,

integration of on-chip QCM with the

electrochemistry technique is realized by fabricating

a semicircular counter electrode next to top electrode

on the same side of the quartz crystal. Variables of d

and w indicate the diameter of working and width of

counter electrode respectively. The gap, g, is the

distance size between working and counter

electrodes.

Figure 1: (a) Design of integrated biosensor. (b) Cross-

section view of integrated biosensor. It indicates the

frequency and impedance changes due to the binding of

bio-molecules (antigens-antibodies) to the surface.

2.2 Resonance Frequency

The resonant frequency of the QCM is determined by

the thickness of quartz (Shi et al., 2016). The highest

resonance frequency is needed to produce the best

sensitivity of this integrated biosensor. This

resonance frequency is induced when acoustic waves

propagate between these two electrodes through the

quartz substrate, as AC signal is applied between the

top and bottom electrodes. The resonant frequency of

the QCM is theoretically given as Eq. (1) (Liu et al.,

2013b):

(1)

Specifically, biosensing in liquids contributes

significant properties of density-viscosity and

acoustic impedance effects to the frequency changes

as follows in equation (2) (Kankare, 2002):

























−

−=

Re1

(2)

Where f

is the resonant frequency of the quartz of the

resonator. Δf is its frequency change due to the layer

on the surface of the resonator. Δm is the change of

mass which is proportional to the change of frequency

and h

is the thickness of the resonator. ρ, ƞ, and µ is

the density, viscosity and shear modulus with indices

l, f, and q indicating the liquid, film (electrode), and

quartz respectively. The material properties and

simulation work used in this work are detailed in

(Zainuddin et al., 2018, 2016).

2.3 Capacitance

Electrochemical biosensors measure current or

impedance changes in electrode surface due to redox

reaction or surface reactions (Wang, 2006).

Generally, these sensors use three-electrode setup for

in situ electrochemistry systems. Firstly, working

electrode is the main active electrode on which the

nano-range changes of chemical reactions occur

(Koyun et al., 2012). Secondly, counter electrode is

used to apply the electric current. Finally, reference

electrode is utilized to provide the reference voltage

for electrochemical tests. During the electro-

migration of ions mobility process, the change of

resistance and diffusion current will be affected on

working electrode surface due to the successful

capture of specific biological targets (Rezaei et al.,

2016). Higher current density is a crucial design

consideration as it corresponds to higher

measurement sensitivity. Apart from that, increasing

the total capacitance or constant phase element (CPE)

in the system level is needed to maximize sensor

sensitivity. All the theoretical studies regarding on

this concept is detailed in (Zainuddin et al., 2016).

The total capacitance or CPE, at electrode-electrolyte

interface denoted as in equation (4) (Nevill and

Malleo, 2012):

CPE

GEO

εε

(3)

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222

Where ε

is the permittivity of free space and has the

value 8.854 x 10

-12

-1

. ε

is the relative dielectric

constant of the medium between two electrodes. A

GEO

is the electrode active geometry surface area. g is the

gap between electrodes. From this equation, reducing

the gap g will increase total capacitance in this

system.

3 PRELIMINARY RESULTS

3.1 COMSOL Simulation

The mechanical (resonance frequency) and

electrochemical operation in the IEQCM array were

simulated using COMSOL 5.3. The detail of

simulations were discussed previously (Zainuddin et

al., 2017).

3.1.1 Resonance Frequency

Figure 2 indicates the measurements of resonance

frequency for the working electrode diameter, from

200 μm to 4000 μm. From the results, the highest

total-displacement value (170 pm) with the resonance

frequency of 9.826 MHz (i.e. natural frequency of

168 μm quartz wafer) was produced at diameter of

4000 μm. Consequently, d = 4000 µm was selected

since it showed the highest total displacement

compared to other diameter based on simulation

results.

Figure 2: Total-displacement versus resonance frequency

when electrode of working diameter, d = 200 μm, 600 μm,

1000 μm, 1400 μm, 1800 μm, 2000 μm, and 4000 μm. The

dotted line box represents the total displacement for all

diameter values around quartz resonance frequency of

9.826 MHz.

3.1.2 Frequency Interference

The optimal centre-to-centre distance of the adjacent

electrodes, (s) was chosen based on the lower

resonance frequency interference. It also produced

the same response of resonance frequency for all

mass sensors. Figure 3 shows the resonance

frequency interference simulation of the centre to

centre distance, (s). From the results, the resonance

frequency interference decreased with the

enlargement of the distance. From the results, s = 4

mm (Figure 3(a)) indicated only two peaks were

observed at the close resonance frequency values. For

s = 6 mm (Figure 3(b)), all QCM sensors showed the

same response resonance frequency of f

= 9.8420

MHz. Finally, for s = 8 mm (Figure 3(c)), it was

observed that, all QCM sensors showed the same

response of resonance frequencies, however the

resonance peaks were not uniform due to electrodes

position near to the edge of quartz. Therefore, s = 6

mm was indicated as the optimal value to allow

minimization of the signal interference between other

channels.

3.1.3 Determination of Gap Separation

The optimal gap between working and counter

electrodes was selected based on the higher

magnitude of total capacitance. Figure 4 shows that

the gap of 70 µm was indicated as the optimal result

to allow maximization of total capacitance in this

simulation as compared to others. Apart from that, it

was observed that widening the counter electrode

width will increase the total capacitance in this

design. Therefore, the gap of (d = 70 µm) and the

counter width of 1000 µm were chosen since these

values showed the highest total capacitance as

compared to other parameters studied in this work.

3.1.4 Fabricated Final Design

Diameter 4000 µm was selected for our biosensor

since it showed an optimal size with a high

mechanical displacement and low interference while

also allowing dimensional fit into a radial array of 3

identical sensors on a single quartz crystal substrate

of diameter, d

= 14 mm as shown in Figure 5. Sensor

was fabricated using standard photolithography

process. Fabrication work was performed in XLIM

Research Institute, University of Limoges, France.

Development of Integrated Electrochemical–Quartz Crystal Microbalance Biosensor Arrays: Towards Ultrasensitive, Multiplexed and Rapid

Point-of-Care Dengue Detection

223

Figure 3: Simulation of frequency interference between the

3 QCM sensors placed symmetrically in the centre of

quartz. (a) s = 4 mm, (b) s = 6 mm, (c) s = 8 mm.

Figure 4: Simulation of total capacitance for gap of 70 μm

to 400 μm and counter width of 200 μm to 1000 μm.

Figure 5: Proposed design and dimension of integrated

electrochemical quartz crystal microbalance in array

(IEQCM) biosensor in a single chip. This sensor has an

array of 3 identical integrated biosensors (IEQCM1,

IEQCM2, IEQCM3) equipped with dual-function detection

system.

3.2 Detection of NS1 Dengue based on

Antigen and Antibody Interactions

In this work, the dengue detection was based on

antigen and antibody binding. Several concentrations

of NS1 antigens (10 ng mL

-1

, 100 ng mL

-1

, 1000 ng

-1

) were measured for demonstration of the

functionality of the IEQCM using a custom

enclosure. Electrochemical impedance spectroscopy

(EIS) and QCM were used to investigate the binding

interactions. Prior to NS1 antigen injection, the

working electrode was coated with self-assembly

monolayer to immobilize the anti-NS1 IgG

antibodies. It was followed by the incubation of

glycine in PBS to block the nonspecific interaction.

Figure 6 shows the measurement results of EIS and

frequency changes before and after incubation with

anti-NS1 reflect the concentrations of NS1. The

electrochemical signals were recorded in the presence

of a redox probe ([Fe(CN)

]

3−/4−

) via potentiostat to

BIODEVICES 2019 - 12th International Conference on Biomedical Electronics and Devices

224

monitor changes in charge-transfer resistance

associated with target binding. The frequency

changes were monitored using OpenQCM software

by injecting 100 µl of phosphate-buffered saline

(PBS) onto the sensor. The frequency change

measurement was started after a stable baseline was

formed as shown in Figure 6(b). Using this new

measuring system, we reliably detected

concentrations of dengue NS1 down to 10 ng mL

-1

well below clinical diagnostic range. Early

measurements of three common human serum

proteins shows that EIS and QCM signals from

nonspecific proteins are no higher than 3 standard

deviations of blank measurements, suggesting the

sensor is highly NS1-selective, however further

investigation is required. Based on these preliminary

results, further validation of the IEQCM will be

performed to allow calibration and to specify the limit

of detection and sensitivity. These validation steps

will support further development of the technology,

and translation of the IEQCM into clinical

applications in the future.

Figure 6: NS1 dengue detection: (a) Nyquist plots from EIS,

(b) frequency change from QCM.

4 SCIENTIFIC AND

SOCIO-ECONOMIC

IMPACTS

From a scientific innovation perspective, we present

the first multichannel array IEQCM biosensor ever

reported. This biosensor brings together

biotechnology, semiconductor and electronics on a

single chip in providing rapid detection of dengue,

noted as the fastest-growing mosquito-borne viral

infection. The array of 3 identical integrated

biosensors equipped with dual-function detection

system offers high-throughput functionalities in

detecting a wide range of dengue antigen

concentrations. Additionally, the ability to multiplex

and parallelize biosensing on a single platform will

allow development of integrated multi-disease

detection panels, such as Aedes-related infections

(dengue, Zika, chikungunya, yellow fever). A

portable, integrated instrument is currently under

active development. Further developments will

enable the device to be operated conveniently by

clinical laboratory personnel or semi-trained staffs, as

multiplexed measurement automation and pre-loaded

protocols are developed in the future. These

approaches have the potential to become PoC

applications that improve standard of care,

particularly in resource-limited settings.

Socio-economically, dengue is a disease that

impacts an estimate of 50 million persons annually

across the globe, heavily concentrated in the tropical

and sub-tropical regions (Special Programme for

Research and Training in Tropical Diseases and

World Health Organization, 2009). By enabling

deployment of an ultrasensitive rapid PoC diagnostic,

a better healthcare delivery for dengue can be

achieved. The portability of the device allows for use

in smaller clinics that often have wider reach into

communities as compared to hospitals with full

laboratory capacity. The prototype is estimated to be

able to run sample analyses in less than 30 minutes.

This rapid analysis allows same-day sample-to-

results cycle, which facilitates faster decisions on

treatment plans for febrile patients suspected of

dengue infection. Perhaps the most novel merit of this

device is the ability to directly detect dengue virus in

solution at very low concentrations, well below

typical clinical diagnostic range of dengue fever,

which promises potential integration into screening

programs for early, pre-viremia detection of dengue

infection. It also could potentially enable screening

for non-asymptomatic carriers for epidemiological

study purposes.

Development of Integrated Electrochemical–Quartz Crystal Microbalance Biosensor Arrays: Towards Ultrasensitive, Multiplexed and Rapid

Point-of-Care Dengue Detection

225

5 CONCLUSIONS

In this position paper, we demonstrate the early

development of an integrated EQCM device for

dengue biosensing. Our working hypothesis is that

using dual-function sensors could not only increase

the sensitivity to very low limits of detection and

increase dynamic range of disease quantification, but

also provide higher diagnostic accuracy through

cross-validation of parallel measurement techniques.

This presents potential capability for new clinical

insights, including early detection of dengue

infection, and identification of asymptomatic carriers.

Furthermore, to the best of our knowledge, this is the

first time a multichannel IEQCM array has been

described. To a larger context, we propose that it is a

worthwhile endeavour to explore more parallel dual-

measurement techniques and their integration

strategies to enhance sensitivity and accuracy of

diagnostic tests.

When completed, this technology could

potentially improve dengue patient care significantly,

as dengue could be detected at an earlier stage, and

enables a faster and higher accuracy of diagnosis at

the point-of-care. As this technology utilizes a

generic immune-affinity sensing scheme, this

technique could also be expanded into other diseases.

ACKNOWLEDGEMENTS

This is a collaborative research between Linkoping

University (Sweden), XLIM Research Institute,

University of Limoges (France) and International

Islamic University Malaysia. It is funded by the

Swedish Research Council (2014-4254) and the

Malaysian Ministry of Higher Education under

FRGS15-217-0458.

REFERENCES

Ahmad, R., Suzilah, I., Wan Najdah, W. M. A., Topek, O.,

Mustafakamal, I., Lee, H. L., 2018. Factors determining

dengue outbreak in Malaysia. PLOS ONE 13, e0193326.

https://doi.org/10.1371/journal.pone.0193 326

Ashshi, A. M., 2017. The prevalence of dengue virus

serotypes in asymptomatic blood donors reveals the

emergence of serotype 4 in Saudi Arabia. Virology

Journal 14. https://doi.org/10.1186/s12985-017-0768-7

Casenghi, M., Kosack, C., Li, R., Bastard, M., Ford, N.,

2014. NS1 antigen detecting assays for diagnosing

acute dengue infection in people living in or returning

from endemic countries. Cochrane Database of

Systematic Reviews.

Cecchetto, J., Carvalho, F. C., Santos, A., Fernandes, F. C.,

Bueno, P.R., 2015. An impedimetric biosensor to test

neat serum for dengue diagnosis. Sensors and Actuators

B: Chemical 213, 150–154.

Darwish, N. T., Alias, Y. B., Khor, S. M., 2015. An

introduction to dengue-disease diagnostics. TrAC

Trends in Analytical Chemistry 67, 45–55.

Dias, A. C. M. S., Gomes-Filho, S. L. R., Silva, M. M. S.,

Dutra, R. F., 2013. A sensor tip based on carbon

nanotube-ink printed electrode for the dengue virus

NS1 protein. Biosensors and Bioelectronics 44, 216–

221. https://doi.org/10.1016/j.bios.2012.12.033

Kankare, J., 2002. Sauerbrey equation of quartz crystal

microbalance in liquid medium. Langmuir 18, 7092–

7094.

Kassim, F. M., Izati, M. N., TgRogayah, T. A., Apandi, Y.

M., Saat, Z., 2011. Use of dengue NS1 antigen for early

diagnosis of dengue virus infection. Southeast Asian J

Trop Med Public Health 42, 562–9.

Koyun, A., Ahlatcolu, E., Koca, Y., Kara, S., 2012.

Biosensors and their principles. A Roadmap of

Biomedical Engineers and Milestones.

Liu, F., Li, F., Nordin, A. N., Voiculescu, I., 2013a. A novel

cell-based hybrid acoustic wave biosensor with

impedimetric sensing capabilities. Sensors 13, 3039–

3055.

Liu, F., Li, F., Nordin, A.N., Voiculescu, I., 2013b. A novel

cell-based hybrid acoustic wave biosensor with

impedimetric sensing capabilities. Sensors 13, 3039–

3055.

Ma, F., Rehman, A., Sims, M., Zeng, X., 2015.

Antimicrobial Susceptibility Assays Based on the

Quantification of Bacterial Lipopolysaccharides via a

Label Free Lectin Biosensor. Anal. Chem. 87, 4385–

4393. https://doi.org/10.1021/acs.analchem.5b00165

Mansor, A.F.M., Ibrahim, S.N., 2016. Simulation of ring

interdigitated electrode for dielectrophoretic trapping,

in: Semiconductor Electronics (ICSE), 2016 IEEE

International Conference On. IEEE, pp. 169–172.

Masrinoul, P., Diata, M.O., Pambudi, S., Limkittikul, K.,

Ikuta, K., Kurosu, T., 2011. Highly conserved region

141–168 of the NS1 protein is a new common epitope

region of dengue virus. Jpn J Infect Dis 64, 109–15.

Mohammed, M., Abakar, A.D., Bakri, Y.M., Dafalla, O.M.,

2018. Molecular surveillance of dengue infections in

Sabya governate of Jazan region, Southwestern Saudi

Arabia.

Nevill, J. T., Malleo, D., 2012. Nanogap Biosensors, in:

Encyclopedia of Nanotechnology. Springer, pp. 1544–

1552.

Omar, N. A. S., Fen, Y. W., 2018. Recent development of

SPR spectroscopy as potential method for diagnosis of

dengue virus E-protein. Sensor Review 38, 106–116.

Pal, S., Dauner, A. L., Valks, A., Forshey, B. M., Long, K.

C., Thaisomboonsuk, B., Sierra, G., Picos, V., Talmage,

S., Morrison, A. C., others, 2015. Multi-country

prospective clinical evaluation of two ELISAs and two

BIODEVICES 2019 - 12th International Conference on Biomedical Electronics and Devices

226

rapid diagnostic tests for diagnosing dengue fever.

Journal of Clinical Microbiology JCM–03042.

Regiart, M., Pereira, S. V., Bertolino, F. A., Garcia, C. D.,

Raba, J., Aranda, P. R., 2016. An electrochemical

immunosensor for anti-T. cruzi IgM antibodies, a

biomarker for congenital Chagas disease, using a

screen-printed electrode modified with gold

nanoparticles and functionalized with shed acute phase

antigen. Microchimica Acta 183, 1203–1210.

Rezaei, B., Ghani, M., Shoushtari, A.M., Rabiee, M., 2016.

Electrochemical biosensors based on nanofibres for

cardiac biomarker detection: A comprehensive review.

Biosensors and Bioelectronics 78, 513–523.

https://doi.org/10.1016/j.bios.2015.11.083

Shafie, A.A., Yeo, H.Y., Coudeville, L., Steinberg, L., Gill,

B.S., Jahis, R., Amar-Singh, H.S.S., 2017. The

Potential Cost Effectiveness of Different Dengue

Vaccination Programmes in Malaysia: A Value-Based

Pricing Assessment Using Dynamic Transmission

Mathematical Modelling. PharmacoEconomics;

Auckland 35, 575–589. http://dx.doi.org/10.1007/

s40273-017-0487-3

Shi, J., Fan, C., Zhao, M., Yang, J., 2016. Energy trapping

of thickness-shear modes in inverted-mesa AT-cut

quartz piezoelectric resonators. Ferroelectrics 494,

157–169.

Silva, E. T., Souto, D. E., Barragan, J. T., Giarola, J. F.,

Moraes, A.C., Kubota, L.T., 2017. Electrochemical

biosensors in point-of-care devices: recent advances

and future trends. ChemElectroChem.

Special Programme for Research and Training in Tropical

Diseases, World Health Organization (Eds.), 2009.

Dengue: guidelines for diagnosis, treatment,

prevention, and control, New ed. ed. TDR : World

Health Organization, Geneva.

Wahyono, T. Y. M., Nealon, J., Beucher, S., Prayitno, A.,

Moureau, A., Nawawi, S., Thabrany, H., Nadjib, M.,

2017. Indonesian dengue burden estimates: review of

evidence by an expert panel. Epidemiology & Infection

145, 2324–2329.

Wang, J., 2006. Analytical electrochemistry. John Wiley &

Sons.

Xiao, C., Zeng, X., 2013. In situ EQCM evaluation of the

reaction between carbon dioxide and electrogenerated

superoxide in ionic liquids. Journal of the

Electrochemical Society 160, H749–H756.

Zainuddin, A. A., Nordin, A. N., Ab Rahim, R., Mak, W.

C., 2016. Modeling of a novel biosensor with integrated

mass and electrochemical sensing capabilities, in:

Biomedical Engineering and Sciences (IECBES), 2016

IEEE EMBS Conference On. IEEE, pp. 420–425.

Zainuddin, A. A., Nordin, A. N., Rahim, R. A., Ralib, A. A.

M., Khan, S., Guines, C., Chatras, M., Pothier, A.,

2018. Verification of Quartz Crystal Microbalance

Array using Vector Network Analyzer and OpenQCM.

Indonesian Journal of Electrical Engineering and

Computer Science 10, 84–93. https://doi.org/10.11591/

ijeecs.v10.i1.pp%p

Zainuddin, A.A., Nordin, A.N., Ralib, A.A.M., Rahim,

R.A., Khan, S., Guines, C., Chatras, M., Pothier, A.,

2017. Design and optimization of a MEMS quartz mass

sensor array for biosensing, in: 2017 IEEE 4th

International Conference on Smart Instrumentation,

Measurement and Application (ICSIMA). Presented at

the 2017 IEEE 4th International Conference on Smart

Instrumentation, Measurement and Application

(ICSIMA), pp. 1–5. https://doi.org/10.1109/ICSIMA.

2017.8311996

Zhang, X., Wang, W., Nordin, A.N., Li, F., Jang, S.,

Voiculescu, I., 2017. The influence of the electrode

dimension on the detection sensitivity of electric cell–

substrate impedance sensing (ECIS) and its

mathematical modeling. Sensors and Actuators B:

Chemical 247, 780–790.

Development of Integrated Electrochemical–Quartz Crystal Microbalance Biosensor Arrays: Towards Ultrasensitive, Multiplexed and Rapid

Point-of-Care Dengue Detection

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