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Rapid multiplex polymerase chain reaction for simultaneous detection of Vibrio harveyi, V. parahaemolyticus, and V. vulnificus in pacific white shrimp (Litopenaeus vannamei)

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dc.contributor.author Thongkao K.
dc.contributor.author Sudjaroen Y.
dc.contributor.author Chaivisuthangkura P.
dc.date.accessioned 2021-04-05T03:23:44Z
dc.date.available 2021-04-05T03:23:44Z
dc.date.issued 2016
dc.identifier.issn 17556783
dc.identifier.other 2-s2.0-84977635826
dc.identifier.uri https://ir.swu.ac.th/jspui/handle/123456789/13407
dc.identifier.uri https://www.scopus.com/inward/record.uri?eid=2-s2.0-84977635826&doi=10.4103%2f1755-6783.184792&partnerID=40&md5=f7d41d6ef1e0261612245eb7e2e9a0a4
dc.description.abstract Context: A comparatively small number of species, e.g., Vibrio parahaemolyticus and V. vulnificus, cause disease in both aquatic animals and humans. V. harveyi is marine animal pathogen and rarely causes infections in humans; however, it might become a reservoir of antibiotic-resistant bacteria forms and virulence genes. Aims: 1) to develop rapid multiplex polymerase chain reaction (PCR) assay for the simultaneous detection of V. harveyi, V. parahaemolyticus, and V. vulnificus by using vhhP2, tl, and rpoS genes as the respective target genes and 2) to evaluate specificity and determined detection of multiplex PCR technique. Materials and Methods: The multiplex PCR assay was developed and evaluated for specificity on 36 isolates of V. harveyi, 30 isolates of V. parahaemolyticus, and 14 isolates of V. vulnificus, along with other species of Vibrio and non-Vibrio bacterial isolates. Sensitivity of test was described as detection limit of pathogens in lowest amount of sample (CFU/mL or CFU/g) was determined by diluted DNA extracts of the pure cultures and spiked pacific white shrimp (Litopenaeus vannamei) samples Results: This developed multiplex PCR was proved as an accurate method, which was specific for three Vibrio species. The detection limits of V. harveyi, V. parahaemolyticus, and V. vulnificus in pure cultures and spiked shrimp samples ranged 1.05-4.8 × 103 CFU/mL and 1.9-7 × 104 CFU/g, respectively. Conclusions: This rapid multiplex PCR assay can decrease amount and process of sample preparation, which was time-consuming, and had preferable accuracy. This developed technique will be suitable and useful for food-borne pathogen detection in shrimp and horizontal gene transfer study among different Vibrio species in aquatic animals.
dc.subject bacterial DNA
dc.subject RNA 16S
dc.subject antibiotic resistance
dc.subject Article
dc.subject bacterial gene
dc.subject bacterium culture
dc.subject bacterium detection
dc.subject bacterium isolate
dc.subject colony forming unit
dc.subject cross reaction
dc.subject DNA extraction
dc.subject DNA template
dc.subject electrophoresis
dc.subject gene targeting
dc.subject horizontal gene transfer
dc.subject limit of detection
dc.subject Litopenaeus vannamei
dc.subject multiplex polymerase chain reaction
dc.subject nonhuman
dc.subject real time polymerase chain reaction
dc.subject Vibrio harveyi
dc.subject Vibrio parahaemolyticus
dc.subject Vibrio vulnificus
dc.title Rapid multiplex polymerase chain reaction for simultaneous detection of Vibrio harveyi, V. parahaemolyticus, and V. vulnificus in pacific white shrimp (Litopenaeus vannamei)
dc.type Article
dc.rights.holder Scopus
dc.identifier.bibliograpycitation Annals of Tropical Medicine and Public Health. Vol 9, No.4 (2016), p.255-262
dc.identifier.doi 10.4103/1755-6783.184792


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