Sandwich Electrochemical Immunosensor for Early Detection of Tuberculosis Based on Graphene/Polyaniline-Modified Screen-Printed Gold Electrode

A rapid and sensitive sandwich electrochemical immunosensor was developed based on the fabrication of the graphene/polyaniline (GP/PANI) nanocomposite onto screen-printed gold electrode (SPGE) for detection of tuberculosis biomarker 10-kDa culture filtrate protein (CFP10). The prepared GP/PANI nanocomposite was characterized using Fourier transform infrared spectroscopy (FTIR) and field emission scanning electron microscopy (FESEM). The chemical bonding and morphology of GP/PANI-modified SPGE were studied by Raman spectroscopy and FESEM coupled with energy dispersive X-ray spectroscopy, respectively. From both studies, it clearly showed that GP/PANI was successfully coated onto SPGE through drop cast technique. Cyclic voltammetry was used to study the electrochemical properties of the modified electrode. The effective surface area for GP/PANI-modified SPGE was enhanced about five times compared with bare SPGE. Differential pulse voltammetry was used to detect the CFP10 antigen. The GP/PANI-modified SPGE that was fortified with sandwich type immunosensor exhibited a wide linear range (20–100 ng/mL) with a low detection limit of 15 ng/mL. This proposed electrochemical immunosensor is sensitive, low sample volume, rapid and disposable, which is suitable for tuberculosis detection in real samples.

[1]  Agustín Costa-García,et al.  Immunosensor for Mycobacterium tuberculosis on screen-printed carbon electrodes. , 2005, Biosensors & bioelectronics.

[2]  Qin Xu,et al.  A paper disk equipped with graphene/polyaniline/Au nanoparticles/glucose oxidase biocomposite modified screen-printed electrode: toward whole blood glucose determination. , 2014, Biosensors & bioelectronics.

[3]  Charles E McCulloch,et al.  Test variability of the QuantiFERON-TB gold in-tube assay in clinical practice. , 2013, American journal of respiratory and critical care medicine.

[4]  U. Hashim,et al.  Enhanced sensing of dengue virus DNA detection using O2 plasma treated-silicon nanowire based electrical biosensor. , 2016, Analytica chimica acta.

[5]  Yang Fan,et al.  Graphene–polyaniline composite film modified electrode for voltammetric determination of 4-aminophenol , 2011 .

[6]  Lan N. Nguyen,et al.  Development of Antigen Detection Assay for Diagnosis of Tuberculosis Using Sputum Samples , 2000, Journal of Clinical Microbiology.

[7]  Yali Shi,et al.  Preparation of alkanethiolate-functionalized core/shell Fe3O4@Au nanoparticles and its interaction with several typical target molecules. , 2008, Analytical chemistry.

[8]  Jaafar Abdullah,et al.  Synthesis and Characterization of Polyaniline/Graphene Composite Nanofiber and Its Application as an Electrochemical DNA Biosensor for the Detection of Mycobacterium tuberculosis , 2017, Sensors.

[9]  Pedro Estrela,et al.  Introduction to biosensors , 2016, Essays in biochemistry.

[10]  G. Ruiz-Palacios,et al.  DETECTION OF MYCOBACTERIAL ANTIGENS IN CEREBROSPINAL FLUID OF PATIENTS WITH TUBERCULOUS MENINGITIS BY ENZYME-LINKED IMMUNOSORBENT ASSAY , 1983, The Lancet.

[11]  Proespichaya Kanatharana,et al.  Ultrasensitive electrochemical immunosensor based on dual signal amplification process for p16(INK4a) cervical cancer detection in clinical samples. , 2015, Biosensors & bioelectronics.

[12]  Kwangnak Koh,et al.  Ultrasensitive immunosensing of tuberculosis CFP-10 based on SPR spectroscopy , 2011 .

[13]  T. Cleary,et al.  Rapid and specific detection of the Mycobacterium tuberculosis complex using fluorogenic probes andreal-time PCR. , 2001, Molecular and cellular probes.

[14]  CheolGi Kim,et al.  Electrochemical biosensor for Mycobacterium tuberculosis DNA detection based on gold nanotubes array electrode platform. , 2016, Biosensors & bioelectronics.

[15]  Keisham Radhapyari,et al.  Graphene-polyaniline nanocomposite based biosensor for detection of antimalarial drug artesunate in pharmaceutical formulation and biological fluids. , 2013, Talanta.

[16]  Linda Zou,et al.  Graphene/Polyaniline nanocomposite as electrode material for membrane capacitive deionization. , 2014 .

[17]  Yohan,et al.  Label-Free Electrochemical DNA Biosensor for the Detection of Mycobacterium Tuberculosis Using Gold Electrode Modified by Self-Assembled Monolayer of Thiol , 2015 .

[18]  Sang-Nae Cho,et al.  Use of rMPB70 Protein and ESAT-6 Peptide as Antigens for Comparison of the Enzyme-Linked Immunosorbent, Immunochromatographic, and Latex Bead Agglutination Assays for Serodiagnosis of Bovine Tuberculosis , 2005, Journal of Clinical Microbiology.

[19]  Jim F Huggett,et al.  Low sensitivity of a urine LAM-ELISA in the diagnosis of pulmonary tuberculosis , 2009, BMC infectious diseases.

[20]  Hafsa Korri-Youssoufi,et al.  Direct E-DNA sensor of Mycobacterium tuberculosis mutant strain based on new nanocomposite transducer (Fc-ac-OMPA/MWCNTs). , 2018, Talanta.

[21]  Tae Jung Park,et al.  Early detection of the growth of Mycobacterium tuberculosis using magnetophoretic immunoassay in liquid culture. , 2017, Biosensors & bioelectronics.

[22]  Jing He,et al.  Dumbbell-like Au-Fe3O4 nanoparticles as label for the preparation of electrochemical immunosensors. , 2010, Biosensors & bioelectronics.

[23]  Uda Hashim,et al.  The utilization of SiNWs/AuNPs-modified indium tin oxide (ITO) in fabrication of electrochemical DNA sensor. , 2014, Materials science & engineering. C, Materials for biological applications.

[24]  Tae Jung Park,et al.  Rapid monitoring of CFP-10 during culture of Mycobacterium tuberculosis by using a magnetophoretic immunoassay , 2013 .

[25]  Xiao-Ming Gao,et al.  Synthesis of orientedly bioconjugated core/shell Fe3O4@Au magnetic nanoparticles for cell separation. , 2011, Talanta.

[26]  A. R. Ruslinda,et al.  Electrical detection of dengue virus (DENV) DNA oligomer using silicon nanowire biosensor with novel molecular gate control. , 2016, Biosensors & bioelectronics.

[27]  Chang Liu,et al.  An electrochemical DNA biosensor for the detection of Mycobacterium tuberculosis, based on signal amplification of graphene and a gold nanoparticle-polyaniline nanocomposite. , 2014, The Analyst.

[28]  Nor Azah Yusof,et al.  Immuno Nanosensor for the Ultrasensitive Naked Eye Detection of Tuberculosis , 2018, Sensors.

[29]  Jonathan P. Metters,et al.  New directions in screen printed electroanalytical sensors: an overview of recent developments. , 2011, The Analyst.

[30]  Marzhan Sypabekova,et al.  Selection, characterization, and application of DNA aptamers for detection of Mycobacterium tuberculosis secreted protein MPT64. , 2017, Tuberculosis.

[31]  A. Pandikumar,et al.  Fabrication of graphene/gold-modified screen-printed electrode for detection of carcinoembryonic antigen. , 2016, Materials science & engineering. C, Materials for biological applications.

[32]  Danila Moscone,et al.  Fully integrated ready-to-use paper-based electrochemical biosensor to detect nerve agents. , 2017, Biosensors & bioelectronics.

[33]  Nor Azah Yusof,et al.  PNA biosensor based on reduced graphene oxide/water soluble quantum dots for the detection of Mycobacterium tuberculosis , 2017 .

[34]  D. A. Russo,et al.  Determination of mycobacterial antigens in sputum by enzyme immunoassay , 1986, Journal of clinical microbiology.

[35]  Jang-Jih Lu,et al.  Detecting Mycobacterium tuberculosis in Bactec MGIT 960 cultures by inhouse IS6110-based PCR assay in routine clinical practice. , 2009, Journal of the Formosan Medical Association = Taiwan yi zhi.

[36]  K. Kaul,et al.  Molecular detection of Mycobacterium tuberculosis: impact on patient care. , 2001, Clinical chemistry.

[37]  Ping Wang,et al.  Screen-printed gold electrode with gold nanoparticles modification for simultaneous electrochemical determination of lead and copper , 2015 .

[38]  V. Choudhry,et al.  Detection of Mycobacterium tuberculosis Antigens in Urinary Proteins of Tuberculosis Patients , 2001, European Journal of Clinical Microbiology and Infectious Diseases.

[39]  M. A. Alonso-Lomillo,et al.  Recent developments in the field of screen-printed electrodes and their related applications. , 2007, Talanta.

[40]  Zhenhua Dai,et al.  A multiple-antigen detection assay for tuberculosis diagnosis based on broadly reactive polyclonal antibodies , 2017, Iranian journal of basic medical sciences.