Rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time PCR

Patricia Rowena, Anak Mark Baran Saging (2012) Rapid detection and quantification of vibrio parahaemolyticus in cockles (anadara granosa) by using real-time PCR. [Final Year Project Report] (Unpublished)

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Abstract

Vibrio parahaemolyticus (V. parahaemolyticus) is acknowledged as one of the most significant disease-causing foodbome pathogen that causes the gastroenteritis resulting in diarrhea from vulnerable patients. The pathogenicity genes of V. parahaemolyticus are thermostable direct hemolysin (tdh) and thermostable direct hemolysin related hemolysin (trh). However, targeting a gene of thermolabile hemolysin (tlh) gene in this study, is described in total V. parahaemolyticus which may play its role in affecting the host and contribute to human health risks. This research aims for V. parahemolyticus in rapidly detecting and quantitatively determines the present and the level of the bacteria from 30 samples of cockles (Anadara granosa) purchased from Kuching7'~-Mile wet market by utilizing the Real-Time PCR. In this study, the primer MV2B-tl (tlh-gene based) and the SYBR Green PCR mastermix were used in Real-Time PCR assay and among all the 30 samples of cockles isolated, there were no detectable quantification of V. parahaemolyticus. Quantification of V. parahaemolyticus could be determined by the standard curve result obtained. Spiking of 'Ti/apia' fish with Ixl07 CFU/mL V. parahaemolyticu aids in determining the sensitivity of the study by Real-Time PCR assay by standard curve plot obtained with ~ value of 0.99 which showed a good correlation. Furthermore, the specificity of the assay was confinned by testing with 7 non -Vibrio strains by specific PCR. The SYBR Green PCR mastermix and primer set were developed as quantification tool of V. parahaemolyticus to the traditional PCR method which needed post-peR result and is time consuming. Therefore, the development of Real-Time peR assay in analyzing the presence of bacteria in seafood is crucial to both seafood industries as well as to the consumers.

Item Type: Final Year Project Report
Additional Information: Project Report (B.Sc.) -- Universiti Malaysia Sarawak, 2012.
Uncontrolled Keywords: Vibrio parahaemolyticus, tlh-gene, Anadara granosa, peR, Real-Time PCR, research, Universiti Malaysia Sarawak, unimas, university, universiti, Borneo, Malaysia, Sarawak, Kuching, Samarahan, ipta, education, undergraduate
Subjects: Q Science > QL Zoology
Divisions: Academic Faculties, Institutes and Centres > Faculty of Resource Science and Technology
Depositing User: Karen Kornalius
Date Deposited: 02 Jul 2015 00:50
Last Modified: 02 Jul 2015 00:50
URI: http://ir.unimas.my/id/eprint/8220

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