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Two Rapid Methods to Identify Three Species of Pathogenic Vibrio in Penaeus vannamei

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Title:Two Rapid Methods to Identify Three Species of Pathogenic Vibrio in Penaeus vannamei
Authors:Luo, K.
Yuan, H.
Zhang, W.
Xu, Q.
Keywords:Penaeus vannamei
Vibrio fluvialis
Vibrio anguillarum
Vibrio alginolyticus
multiplex PCR
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LC Subject Headings:Fish culture--Israel.
Fish culture
Date Issued:2018
Abstract:Vibrio species frequently infect Penaeus vannamei causing great economic losses to the
Whiteleg shrimp industry. Rapid detection of pathogenic Vibrio infection would improve
the fight against these diseases. In this study, single and multiple polymerase chain
reaction (PCR) methods were developed to detect three species of pathogenic Vibrio:
Vibrio fluvialis, Vibrio anguillarum, and Vibrio alginolyticus. Specific primers were
designed for the toxR gene of V. fluvialis, the flaA gene of V. anguillarum, and the pyrH
gene of V. alginolyticus. The bacteria were used as templates to establish a 25 μL
reaction system for PCR amplification. The results showed that single and specific PCR
amplification products of expected sizes were obtained (228bp, 1665bp, and 383bp,
respectively). The lowest concentration detected for the three Vibrio species were
, 2.70×104
, and 2.48×102 colony forming units (cfu)/mL, respectively. We also
developed a multiplex PCR method to identify the three Vibrio species accurately, and
with improved identification efficiency. In addition, quantitative real-time PCR (qPCR)
was used to identify the minimum detectable DNA concentration for the three Vibrio
species (1.0×10-6 nmol/L for V. fluvialis, 1.0×10-7 nmol/L for V. anguillarum, and 1.0×10-
8 nmol/L for V. alginolyticus). Technical requirements for ordinary PCR are low, therefore
PCR is a feasible technique to detect and diagnose Penaeus vannamei bacterial disease.
Pages/Duration:7 pages
Appears in Collections: Volume 70, 2018

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