A multiplex PCR-based protocol for identification and quantification of Vibrio harveyi-related species

Cano-Gomez, Ana, Høj, Lone, Owens, Leigh, Baillie, Brett K., and Andreakis, Nikos (2015) A multiplex PCR-based protocol for identification and quantification of Vibrio harveyi-related species. Aquaculture, 437. pp. 195-200.

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Abstract

Vibrios of the Harveyi clade are responsible for severe economic losses in the aquaculture industry worldwide, but so far no rapid method has been available to discriminate between the closely related species Vibrio campbellii, Vibrio harveyi, Vibrio owensii and Vibrio rotiferianus. Here, we describe a multiplex PCR assay capable of fast and reliable detection and identification of these species by amplification of three conserved protein-coding genes (topA, ftsZ and mreB). The developed species-specific primer sets produce PCR amplicons of unequal length and can be used separately or in combination. When combined with a 3-tube most-probable-number (MPN) based protocol, the multiplex PCR allows highly sensitive detection and enumeration in complex, aquaculture-relevant samples such as water, Artemia and spiny lobster (Panulirus ornatus) larvae. The described multiplex PCR represents a significant advance for aquaculture by providing an easy, fast and low-cost method for identification of several closely related pathogens that are difficult to discriminate by traditional approaches.

Item ID: 40658
Item Type: Article (Research - C1)
ISSN: 1873-5622
Keywords: Vibrio harveyi; Vibrio campbellii; Vibrio rotiferianus; Vibrio owensii; Aquaculture; Multiplex PCR-based identification protocol
Funders: James Cook University (JCU), AIMS@JCU
Projects and Grants: AIMS@JCU bursary No. 6281.98923.2690
Date Deposited: 06 Oct 2015 01:40
FoR Codes: 06 BIOLOGICAL SCIENCES > 0605 Microbiology > 060501 Bacteriology @ 50%
07 AGRICULTURAL AND VETERINARY SCIENCES > 0704 Fisheries Sciences > 070404 Fish Pests and Diseases @ 50%
SEO Codes: 83 ANIMAL PRODUCTION AND ANIMAL PRIMARY PRODUCTS > 8301 Fisheries - Aquaculture > 830101 Aquaculture Crustaceans (excl. Rock Lobster and Prawns) @ 100%
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