NDLI: Piezoresistive microcantilever-based DNA sensor for sensitive detection of pathogenic Vibrio cholerae O1 in food sample.

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Piezoresistive microcantilever-based DNA sensor for sensitive detection of pathogenic Vibrio cholerae O1 in food sample.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Suppat, Assawapong Khemthongcharoen, Numfon Jaruwongrungsee, Kata Tuantranont, Adisorn Wonglumsom, Wijit Promptmas, Chamras
Description	Country affiliation: Thailand Author Affiliation: Khemthongcharoen N ( Department of Clinical Chemistry, Faculty of Medical Technology, Mahidol University, 999 Phuttamonthon 4 Rd., Salaya, Nakhon Pathom 73170, Thailand); Wonglumsom W ( Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand.); Suppat A ( Nanoelectronics and MEMS Laboratory, National Electronics and Computer Technology Center, Pathumthani, Thailand.); Jaruwongrungsee K ( Nanoelectronics and MEMS Laboratory, National Electronics and Computer Technology Center, Pathumthani, Thailand.); Tuantranont A ( Nanoelectronics and MEMS Laboratory, National Electronics and Computer Technology Center, Pathumthani, Thailand.); Promptmas C ( Department of Clinical Chemistry, Faculty of Medical Technology, Mahidol University, 999 Phuttamonthon 4 Rd., Salaya, Nakhon Pathom 73170, Thailand. Electronic address: chamras.pro@mahidol.ac.th.)
Abstract	Pathogenic Vibrio cholerae produces a cholera toxin which is the cause of a severe diarrheal disease called 'Cholera'. Available detection methods, including standard bacteriological test and immuno-based detection, are specific to the suspected pathogenic V. cholerae O1 and O139, but they are not specific to the cholera toxin producible strain. This work combined the polymerase chain reaction (PCR) of cholera toxin gene, ctxA gene, and microcantilever-based DNA sensor to improve the sensitivity and specificity of detection. Gold coated microcantilever, 250 µm long and 50 µm wide, with an embedded polysilicon wire acting as a piezoresistive material was modified by a self-assembled monolayer (SAM) of 3-mercaptopropionic acid (MPA) for immobilization of specific DNA probe via avidin layer on the surface. The avidin and 5' biotinylated single-stranded DNA (ssDNA) probe concentrations were optimized for the immobilization at 50 µg/mL and 1 µM, respectively. The hybridization between ssDNA probe on this DNA sensor and target DNA creates nanomechanical bending and resistance change of piezoresistive material inside the beam. This microcantilever-based DNA sensor offers a detection sensitivity of 3.25 pg or 14 nM of DNA template for ctxA gene detection. The lowest number of V. cholerae O1 in food sample with and without the enrichment process that the polymerase chain reaction (PCR) for ctxA gene combined with this DNA sensor can detect is 0.835 and 835 cells/g, respectively. This detection sensitivity is 10 times higher than that of the conventional PCR method.
ISSN	09565663
Volume Number	63
e-ISSN	18734235
Journal	Biosensors and Bioelectronics
Language	English
Publisher	Elsevier
Publisher Date	2015-01-15
Publisher Place	Great Britain (UK)
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Biosensing Techniques Methods Cholera Diagnosis Dna, Bacterial Isolation & Purification Vibrio Cholerae O1 Microbiology Cholera Toxin Chemistry Foodborne Diseases Humans Genetics Pathogenicity Journal Article Research Support, Non-u.s. Gov't Discipline Biotechnology
Content Type	Text
Resource Type	Article
Subject	Nanoscience and Nanotechnology Medicine Biophysics Biomedical Engineering Biotechnology Electrochemistry

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