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Electrochemical genosensor for specific detection of the food-borne pathogen, Vibrio cholerae

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Abstract

A disposable horseradish peroxidase (HRP)-based electrochemical genosensor was developed for chronoamperometric detection of single-stranded asymmetric lolB gene PCR amplicon (118 bp in length) of the food-borne pathogen, Vibrio cholerae. A two-step sandwich-type hybridization strategy using two specific probes was employed for specific detection of the target single-stranded DNA (ssDNA). The analytical performances of the detection platform have been evaluated using a synthetic ssDNA (ST3) which was identical to the target single-stranded amplicon and a total of 19 bacterial strains. Under optimal condition, ST3 was calibrated with a dynamic range of 0.4883–15.6250 nM. By coupling asymmetric PCR amplification, the probe-based electrochemical genosensor was highly specific to the target organism (100% specificity) and able to detect as little as 0.85 ng/μl of V. cholerae genomic DNA.

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References

  • Bakker E (2004) Electrochemical sensors. Anal Chem 76(12):3285–3298

    Article  CAS  Google Scholar 

  • Bekaert M, Teeling EC (2008) UniPrime: a workflow-based platform for improved universal primer design. Nucleic Acids Res 36(10):e56

    Article  Google Scholar 

  • Bopp CA, Ries AA, Well JG (1999) Laboratory methods for the diagnosis of epidemic dysentery and cholera. Centers for disease control and prevention

  • Bunyakul N, Edwards K, Promptmas C, Baeumner A (2009) Cholera toxin subunit B detection in microfluidic devices. Anal Bioanal Chem 393(1):177–186

    Article  CAS  Google Scholar 

  • Carter RM, Mekalanos JJ, Jacobs MB, Lubrano GJ, Guilbault GG (1995) Quartz crystal microbalance detection of Vibrio cholerae O139 serotype. J Immunol Methods 187(1):121–125

    Article  CAS  Google Scholar 

  • Charles RC, Ryan ET (2011) Cholera in the 21st century. Curr Opin Infect Dis 24(5):472–477

    Article  Google Scholar 

  • Choopun N, Louis V, Huq A, Colwell RR (2002) Simple procedure for rapid identification of Vibrio cholerae from the aquatic environment. Appl Environ Microbiol 68(2):995–998

    Article  CAS  Google Scholar 

  • Del Giallo ML, Ariksoysal DO, Marrazza G, Mascini M, Ozsoz M (2005) Disposable electrochemical enzyme-amplified genosensor for Salmonella bacteria detection. Anal Lett 38(15):2509–2523

    Article  Google Scholar 

  • Dieffenbach CW, Lowe TMJ, Dveksler GS (1993) General concepts for PCR primer design. Genome Res 3(3):S30–S37

    Article  CAS  Google Scholar 

  • Djellouli N, Rochelet-Dequaire M, Limoges B, Druet M, Brossier P (2007) Evaluation of the analytical performances of avidin-modified carbon sensors based on a mediated horseradish peroxidase enzyme label and their application to the amperometric detection of nucleic acids. Biosens Bioelectron 22(12):2906–2913

    Article  CAS  Google Scholar 

  • Drummond TG, Hill MG, Barton JK (2003) Electrochemical DNA sensors. Nat Biotechnol 21(10):1192–1199

    Article  CAS  Google Scholar 

  • Dudak FC, Boyaci IH (2009) Rapid and label-free bacteria detection by surface plasmon resonance (SPR) biosensors. Biotechnol J 4(7):1003–1011

    Article  CAS  Google Scholar 

  • Ghose AC (2011) Lessons from cholera & Vibrio cholerae. Indian J Med Res 133(2):164–170

    Google Scholar 

  • Goel AK, Jain M, Kumar P, Jiang SC (2010) Molecular characterization of Vibrio cholerae outbreak strains with altered El Tor biotype from southern India. World J Microbiol Biotechnol 26(2):281–287

    Article  CAS  Google Scholar 

  • Hasan JAK, Bernstein D, Huq A, Loomis L, Tamplin ML, Colwell RR (1994) Cholera DFA: an improved direct fluorescent monoclonal antibody staining kit for rapid detection and enumeration of Vibrio cholerae O1. FEMS Microbiol Lett 120(1–2):143–148

    Article  CAS  Google Scholar 

  • Huq A, Colwell RR, Rahman R, Ali A, Chowdhury MA, Parveen S, Sack DA, Russek-Cohen E (1990) Detection of Vibrio cholerae O1 in the aquatic environment by fluorescent-monoclonal antibody and culture methods. Appl Environ Microbiol 56(8):2370–2373

    CAS  Google Scholar 

  • Jubete E, Loaiza OA, Ochoteco E, Pomposo JA, Grande H, RodrÃ-guez J (2009) Nanotechnology: a tool for improved performance on electrochemical screen-printed (bio)sensors. J Sens 2009:1–13

    Article  Google Scholar 

  • Jyoung JY, Hong S, Lee W, Choi JW (2006) Immunosensor for the detection of Vibrio cholerae O1 using surface plasmon resonance. Biosens Bioelectron 21(12):2315–2319

    Article  CAS  Google Scholar 

  • Kerman K, Vestergaard M, Nagatani N, Takamura Y, Tamiya E (2006) Electrochemical genosensor based on peptide nucleic acid-mediated PCR and asymmetric PCR techniques: electrostatic interactions with a metal cation. Anal Chem 78(7):2182–2189

    Article  CAS  Google Scholar 

  • Kwok S, Kellogg DE, McKinney N, Spasic D, Goda L, Levenson C, Sninsky JJ (1990) Effects of primer-template mismatches on the polymerase chain reaction: human immunodeficiency virus type 1 model studies. Nucleic Acids Res 18(4):999–1005

    Article  CAS  Google Scholar 

  • Lai RY, Lagally ET, Lee SH, Soh HT, Plaxco KW, Heeger AJ (2006) Rapid, sequence-specific detection of unpurified PCR amplicons via a reusable, electrochemical sensor. Proc Natl Acad Sci USA 103(11):4017–4021

    Article  CAS  Google Scholar 

  • Lalitha P, Siti Suraiya MN, Lim KL, Lee SY, Nur Haslindawaty AR, Chan YY, Ismail A, Zainuddin ZF, Ravichandran M (2008) Analysis of lolB gene sequence and its use in the development of a PCR assay for the detection of Vibrio cholerae. J Microbial Methods 75(1):142–144

    Article  CAS  Google Scholar 

  • Liu G, Wan Y, Gau V, Zhang J, Wang L, Song S, Fan C (2008) An enzyme-based E-DNA sensor for sequence-specific fetection of femtomolar DNA targets. J Am Chem Soc 130(21):6820–6825

    Article  CAS  Google Scholar 

  • Loyprasert S, Hedström M, Thavarungkul P, Kanatharana P, Mattiasson B (2010) Sub-attomolar detection of cholera toxin using a label-free capacitive immunosensor. Biosens Bioelectron 25(8):1977–1983

    Article  CAS  Google Scholar 

  • Madoroba E, Momba MNB (2010) Prevalence of Vibrio cholerae in rivers of mpumalanga province, south africa as revealed by polyphasic characterization. Afr J Biotechnol 9(43):7295–7301

    CAS  Google Scholar 

  • Martins MT, Sanchez PS, Sato MIZ, Brayton PR, Colwell RR (1993) Detection of Vibrio cholerae O1 in the aquatic environment in Brazil employing direct immunofluorescence microscopy. World J Microbiol Biotechnol 9(3):390–392

    Article  Google Scholar 

  • Mix M, Reske T, Duwensee H, Flechsig G-U (2009) Electrochemical detection of asymmetric PCR products by labeling with osmium tetroxide. Electroanal 21(7):826–830

    CAS  Google Scholar 

  • Navaneethan U, Giannella RA (2008) Mechanisms of infectious diarrhea. Nat Clin Pract Gastroenterol Hepatol 5(11):637–647

    Article  Google Scholar 

  • Pedrero M, Campuzano S, Pingarron JM (2011) Electrochemical genosensors based on PCR strategies for microorganisms detection and quantification. Anal Methods 3(4):780–789

    Article  CAS  Google Scholar 

  • Peressutti SR, Jurquiza V, GonzÁLez-Fraga S, Pichel M, Binsztein N, Costagliola M (2009) Direct fluorescent antibody-direct viable count and polymerase chain reaction detection limit for the identification of Vibrio cholerae O1 in mussels (Mytilus edulis). J Rapid Methods Autom Microbiol 17(3):291–303

    Article  CAS  Google Scholar 

  • Rao VK, Sharma MK, Goel AK, Singh L, Sekhar K (2006) Amperometric immunosensor for the detection of Vibrio cholerae O1 using disposable screen-printed electrodes. Anal Sci 22(9):1207–1211

    Article  CAS  Google Scholar 

  • Sack DA, Sack RB, Nair GB, Siddique AK (2004) Cholera. Lancet 363(9404):223–233

    Article  CAS  Google Scholar 

  • Sharma MK, Goel AK, Singh L, Rao VK (2006) Immunological biosensor for detection of Vibrio cholerae O1 in environmental water samples. World J Microbiol Biotechnol 22(11):1155–1159

    Article  CAS  Google Scholar 

  • Sungkanak U, Sappat A, Wisitsoraat A, Promptmas C, Tuantranont A (2010) Ultrasensitive detection of Vibrio cholerae O1 using microcantilever-based biosensor with dynamic force microscopy. Biosens Bioelectron 26(2):784–789

    Article  CAS  Google Scholar 

  • Tamrakar A, Goel A, Kamboj D, Singh L (2006) Surveillance methodology for Vibrio cholerae in environmental samples. Int J Environ Health Res 16(4):305–312

    Article  Google Scholar 

  • Thomas S, Ramachandran D, Remani B, Singh DV (2008) Detection and molecular characterization of Vibrio cholerae O1 Inaba biotype El Tor strain in Kerala, S. India. World J Microbiol Biotechnol 24:433–434

    Article  CAS  Google Scholar 

  • Tosar JP, Bra˜nas G, Laíz J (2010) Electrochemical DNA hybridization sensors applied to real and complex biological samples. Biosens Bioelectron 26(4):1205–1217

    Article  CAS  Google Scholar 

  • Viswanathan S, Wu LC, Huang MR, Ho JAA (2006) Electrochemical immunosensor for cholera toxin using liposomes and poly(3, 4-ethylenedioxythiophene)-coated carbon nanotubes. Anal Chem 78(4):1115–1121

    Article  CAS  Google Scholar 

  • WHO (2009) Cholera: global surveillance summary, 2008. Wkly Epidemiol Rec 84(31):309–324

    Google Scholar 

  • WHO (2011) Cholera, 2010 Weekly epidemiological record/Health Section of the Secretariat of the League of Nations 86(31):325–339

  • Yean CY, Kamarudin B, Ozkan DA, Yin LS, Lalitha P, Ismail A, Ozsoz M, Ravichandran M (2008) Enzyme-linked amperometric electrochemical genosensor assay for the detection of PCR amplicons on a streptavidin-treated screen-printed carbon electrode. Anal Chem 80(8):2774–2779

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors thank the research funding support received in the form of a Research University (RU) grant (grant no: 1001/PPSP/813020) and a Postgraduate Research Grant Scheme (PRGS) (grant no: 1001/PPSP/8144004) from Universiti Sains Malaysia (USM), Malaysia. In addition, support from the Institute of Postgraduate Studies (IPS), USM in the form of a graduate fellowship to Kim-Fatt Low is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia

    Kim-Fatt Low & Yean-Yean Chan

  2. School of Bioresources and Technology, and Biological Engineering Program, King Mongkut’s University of Technology Thonburi, Bangkhuntien, Bangkok, 10150, Thailand

    Kritsanaporn Chuenrangsikul, Patsamon Rijiravanich & Werasak Surareungchai

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  1. Kim-Fatt Low
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  2. Kritsanaporn Chuenrangsikul
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  3. Patsamon Rijiravanich
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  4. Werasak Surareungchai
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  5. Yean-Yean Chan
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Correspondence to Yean-Yean Chan.

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Low, KF., Chuenrangsikul, K., Rijiravanich, P. et al. Electrochemical genosensor for specific detection of the food-borne pathogen, Vibrio cholerae . World J Microbiol Biotechnol 28, 1699–1706 (2012). https://doi.org/10.1007/s11274-011-0978-x

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  • DOI: https://doi.org/10.1007/s11274-011-0978-x

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