Multiplex MPN-PCR for the enumeration of three major Vibrios in raw fishes in Malaysia

Bonny, Sharmin Quazi and Hossain, M.A. Motalib and Thong, Kwai Lin and Ali, Md. Eaqub (2018) Multiplex MPN-PCR for the enumeration of three major Vibrios in raw fishes in Malaysia. Food Control, 90. pp. 459-465. ISSN 0956-7135

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Official URL: https://doi.org/10.1016/j.foodcont.2018.02.034

Abstract

Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus are the most notorious fish borne-pathogens responsible for several deadly diseases in human. However, the prevalence of these three pathogenic vibrios in Malaysian fishes has not been thoroughly investigated, leaving a study gap for the tracing of possible Vibrio outbreak through carrier fish consumption. The most-probable-number-PCR (MPN-PCR) assays are highly sensitive for the identification of a single bacterial pathogen but they have not been applied for the discrimination of major vibrios in a multiplex platform. This study coupled MPN method with Multiplex PCR to differentiate three major vibrios in a one test platform. A total of 120 fish samples were analyzed and the prevalence of V. parahaemolyticus, V. cholerae, and V. vulnificus was 48.33%, 27%, and 32.50%, respectively. When simplex MPN-PCR was applied similar results were obtained at P < 0.05, validating the accuracy of multiplex MPN-PCR. The microbial loads of V. parahaemolyticus, V. cholerae, and V. vulnificus in the tested samples were 0–2.43 × 106, 0–2.16 × 106, and 0–4.93 × 105 MPN g−1, respectively. Primers based on housekeeping pntA gene amplified V. parahaemolyticus (409 bp) and V. cholerae (338 bp) but virulent vvhA gene detected V. vulnificus (205 bp). Thus the 27–48% prevalence of targeted vibrios in consumable fishes indicated there is considerable risk of Vibrio onset in Malaysia due to the consumption of undercooked or raw fish products.

Item Type: Article
Uncontrolled Keywords: Multiplex MPN-PCR assay; Fish-borne pathogens; Zoonotic vibrios; Microbial load; Raw and undercooked fish products
Subjects: Q Science > Q Science (General)
Q Science > QH Natural history
Divisions: Faculty of Science > Institute of Biological Sciences
Deputy Vice Chancellor (Research & Innovation) Office > Centre for Research in Biotechnology for Agriculture
Deputy Vice Chancellor (Research & Innovation) Office > Nanotechnology & Catalysis Research Centre
Depositing User: Ms. Juhaida Abd Rahim
Date Deposited: 11 Sep 2019 09:04
Last Modified: 11 Sep 2019 09:04
URI: http://eprints.um.edu.my/id/eprint/22302

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