Development of a SYBR green based real-time polymerase chain reaction assay for specific detection and quantification of Vibrio parahaemolyticus from food and environmental samples.

V, Micky and AT Nur, Quraitu and L, Velnetti and RMB, Patricia and C, Christy and MB, Lesley (2014) Development of a SYBR green based real-time polymerase chain reaction assay for specific detection and quantification of Vibrio parahaemolyticus from food and environmental samples. International Food Research Journal, 21 (3). pp. 921-927. ISSN 2231-7546

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Official URL: http://www.ifrj.upm.edu.my/21%20%2803%29%202014/11...

Abstract

Vibrio parahaemolyticus is a foodborne pathogen and their human infection is regularly associated with the consumption of raw or undercooked seafood and contaminated water supplies. Many conventional biochemical identification and confirmation procedures are performed to detect the presence of this pathogen, both from seafood or environmental samples. However, these procedures not only require two or more days to complete, they do not have the capabilities to determine the number of V. parahaemolyticus cells in any given samples. Thus, in this study we describe the development of a rapid SYBR green based real-time PCR assay, targeting the thermo labile (tl) gene of V. parahaemolyticus for the detection and enumeration of this bacterium from seafood and environmental samples. We report that the real-time PCR assay and the primers designed are highly specific, and only generated the desired amplicons with V. parahaemolyticus DNA samples against other bacteria and fungi species. Our assay is also highly sensitive, and, is able to detect V. parahaemolyticus with high coefficient values in concentrations as low as 1.0 pg/μl DNA for pure genomic DNA solutions and 10 cells/ml in serially diluted cell suspension and spiked samples. This assay can be completed in less than 3 hours and may be used as a tool for rapid determination of V. parahaemolyticus densities in the food industries, environmental risk assessment and for clinical diagnostics purposes.

Item Type: Article
Uncontrolled Keywords: Real-time Polymerase Chain Reaction (real-time PCR) , SYBR Green , Thermo labile (tl) gene , Vibrio parahaemolyticus , unimas, university, universiti, Borneo, Malaysia, Sarawak, Kuching, Samarahan, ipta, education ,undergraduate,postgraduate, research, Universiti Malaysia Sarawak.
Subjects: G Geography. Anthropology. Recreation > GE Environmental Sciences
Q Science > QR Microbiology
Divisions: Academic Faculties, Institutes and Centres > Faculty of Resource Science and Technology
Depositing User: Karen Kornalius
Date Deposited: 25 May 2015 08:20
Last Modified: 14 Sep 2015 02:57
URI: http://ir.unimas.my/id/eprint/7387

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