Separation of endogenous viral elements from infectious Penaeus stylirostris densovirus using recombinase polymerase amplification

Jaroenram, Wansadaj, and Owens, Leigh (2014) Separation of endogenous viral elements from infectious Penaeus stylirostris densovirus using recombinase polymerase amplification. Molecular and Cellular Probes, 28 (5-6). pp. 284-287.

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Abstract

Non-infectious Penaeus stylirostris densovirus (PstDV)-related sequences in the shrimp genome cause false positive results with current PCR protocols. Here, we examined and mapped PstDV insertion profile in the genome of Australian Penaeus monodon. A DNA sequence which is likely to represent infectious PstDV was also identified and used as a target sequence for recombinase polymerase amplification (RPA)-based approach, developed for specifically detecting PstDV. The RPA protocol at 37 °C for 30 min showed no cross-reaction with other shrimp viruses, and was 10 times more sensitive than the 309F/R PCR protocol currently recommended by the World Organization for Animal Health (OIE) for PstDV diagnosis. These features, together with the simplicity of the protocol, requiring only a heating block for the reaction, offer opportunities for rapid and efficient detection of PstDV.

Item ID: 37094
Item Type: Article (Research - C1)
ISSN: 1096-1194
Keywords: Penaeus stylirostris; PstDV; PstDV-related sequences; RPA
Funders: James Cook University (JCU), Indo Australia Biotechnology
Projects and Grants: JCU (6281.93551.0208), Indo Australia Biotechnology (BF50090)
Date Deposited: 25 Feb 2015 22:01
FoR Codes: 06 BIOLOGICAL SCIENCES > 0605 Microbiology > 060506 Virology @ 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 > 830105 Aquaculture Prawns @ 100%
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