Development of a sybr green based real-time polymerase chain reaction assay for specific detection and quantification of vibrio parahaemolyticus from selected seafood Key words: Vibrio parahaemolyticus, real-time polymerase chain reaction (RT-PCR), SYBR Green dye, seafood

• Main Author: Nur Quraitu' Aini, Binti Tajudin
• Format: Final Year Project Report
• Language: English
• Published: Universiti Malaysia Sarawak, (UNIMAS) 2012
• Subjects: GE Environmental Sciences; Q Science (General)
• Online Access: http://ir.unimas.my/id/eprint/7363/3/DEVELOPMENT%20OF%20A%20SYBR%20GREEN%20BASED%20REAL-TIME%20POLYMERASE%20CHAIN%20REACTION%20ASSAY%20%28OCR%29.pdf; http://ir.unimas.my/id/eprint/7363/
• Institution: Universiti Malaysia Sarawak

Vibrio parahaemolyticus is a foodborne pathogen and their human infection is regularly associated with the consumption of contaminated, raw or undercooked seafood. To date, many biochemical identification and confirmation are done to detect the presence of this pathogen, especially from seafood samples. However these procedure require 2 or more days to complete. Thus, in this study we describe the development of a rapid SYBR green based real-time PCR assay for the detection and nenumeration of V.parahaemolyticus from seafood sample. The real-time PCR assay and the primers designed were highly specific, and only generated the desired amplicons with V. parahaemolyticus DNA samples against other bacteria and fungi species. Our assay was able to detect V. parahaemolyticus with high coefficient values in concentrations as low as 5.0 pg/μl DNA in pure genomic DNA solutions and 50 CFU/ml in spiked samples.