Detection and quantification of pathogenic Vibrio parahaemolyticus in shellfish by using multiplex PCR and loop-mediated isothermal amplification assay

Malcom, Tan Turk Hsern and Cheah, Yoke Kqueen and Mohamad Radzi, Che Wan Jasimah Wan and Kasim, Faridah Abu and Kantilal, Haresh Kumar and Tang, John Yew Huat and Martinez-Urtaza, Jaime and Nakaguchi, Yoshitsugu and Nishibuchi, Mitsuaki and Radu, Son (2015) Detection and quantification of pathogenic Vibrio parahaemolyticus in shellfish by using multiplex PCR and loop-mediated isothermal amplification assay. Food Control, 47 (2015). pp. 664-671. ISSN 0956-7135

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Abstract

Vibrio parahaemolyticusis a halophilic bacterium that commonly inhabits the marine and estuarine environments. This organism is also one of the leading causative pathogen of gastroenteritis often related to consumption of raw or undercooked seafood. In this study, molluscan shellfish (bloody clams and surf clams) and crustaceans (shrimps) were monitored in wet markets and hypermarkets. Two molecular methods were employed and compared to detect total and pathogenicV. parahaemolyticusin MPN enrichments: multiplex PCR and LAMP assay. The multiplex PCR was optimized to detect the total (toxRþ), tdhþ and trhþV. parahaemolyticus. On the other hand, the LAMP assay was employed to target the pathogenic strains only, the tdhþ and trhþ, respectively. Out of 232 samples examined, 229 (98.7%) were positive for V. parahaemolyticus with counts ranging from 30 to>110, 000 MPN/g. Positive samples for tdhþ V. parahaemolyticus were obtained in 77 out of 232 (33.1%) samples ranging from 30 to>110, 000 MPN/g. Meanwhile, positive samples fortrhþwere identified in 16 out of 232 (6.9%) samples examined ranging from 30 to 9600 MPN/g. Detection of samples with presence oftdhþgenes did not vary between methods, but a significant difference was observed when the LAMP assay was compared to PCR to detect trhþ V. parahaemolyticus. Therefore, on occasions where the density of the targeted genes is low, the LAMP assay serves as a better alternative. Nonetheless, this study constitutes an assessment of presence of total and potentially pathogenicV. parahaemolyticusin shellfishes for domestic consumption revealing the potential risk of contracting vibriosis if precautions and safety measures are not properly managed.

Item Type: Article
Keywords: Vibrio parahaemolyticus: total,tdh and trh, Molluscan shellfish (bloody clams and surf clams), Crustacean shellfish (shrimps), Most probable number (MPN), Multiplex Polymerase Chain Reaction (multiplex PCR), Loop-mediated isothermal amplification (LAMP)
Subjects: S Agriculture > S Agriculture (General)
Faculty / Institute: Faculty of Bioresources & Food Industry
Depositing User: Dr John Yew Huat Tang
Date Deposited: 21 Jan 2015 02:41
Last Modified: 21 Jan 2015 02:41
URI: http://erep.unisza.edu.my/id/eprint/1868

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