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

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...

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Main Authors: Bonny, Sharmin Quazi, Hossain, M.A. Motalib, Thong, Kwai Lin, Ali, Md. Eaqub
Format: Article
Published: Elsevier 2018
Subjects:
Q Science (General)
QH Natural history
Online Access:http://eprints.um.edu.my/22302/
https://doi.org/10.1016/j.foodcont.2018.02.034
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spelling my.um.eprints.223022019-09-11T09:04:52Z http://eprints.um.edu.my/22302/ Multiplex MPN-PCR for the enumeration of three major Vibrios in raw fishes in Malaysia Bonny, Sharmin Quazi Hossain, M.A. Motalib Thong, Kwai Lin Ali, Md. Eaqub Q Science (General) QH Natural history 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. Elsevier 2018 Article PeerReviewed 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 https://doi.org/10.1016/j.foodcont.2018.02.034 doi:10.1016/j.foodcont.2018.02.034
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
topic Q Science (General)
QH Natural history
spellingShingle Q Science (General)
QH Natural history
Bonny, Sharmin Quazi
Hossain, M.A. Motalib
Thong, Kwai Lin
Ali, Md. Eaqub
Multiplex MPN-PCR for the enumeration of three major Vibrios in raw fishes in Malaysia
description 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.
format Article
author Bonny, Sharmin Quazi
Hossain, M.A. Motalib
Thong, Kwai Lin
Ali, Md. Eaqub
author_facet Bonny, Sharmin Quazi
Hossain, M.A. Motalib
Thong, Kwai Lin
Ali, Md. Eaqub
author_sort Bonny, Sharmin Quazi
title Multiplex MPN-PCR for the enumeration of three major Vibrios in raw fishes in Malaysia
title_short Multiplex MPN-PCR for the enumeration of three major Vibrios in raw fishes in Malaysia
title_full Multiplex MPN-PCR for the enumeration of three major Vibrios in raw fishes in Malaysia
title_fullStr Multiplex MPN-PCR for the enumeration of three major Vibrios in raw fishes in Malaysia
title_full_unstemmed Multiplex MPN-PCR for the enumeration of three major Vibrios in raw fishes in Malaysia
title_sort multiplex mpn-pcr for the enumeration of three major vibrios in raw fishes in malaysia
publisher Elsevier
publishDate 2018
url http://eprints.um.edu.my/22302/
https://doi.org/10.1016/j.foodcont.2018.02.034
_version_ 1646210202321551360
score 13.211869

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