title: Separation of endogenous viral elements from infectious Penaeus stylirostris densovirus using recombinase polymerase amplification.
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.
authors.affiliation: School of Veterinary and Biomedical Sciences, James Cook University, Townsville, Queensland, 4811, Australia.
authors.forename: Wansadaj
authors.initials: W
authors.lastname: Jaroenram
authors.affiliation: School of Veterinary and Biomedical Sciences, James Cook University, Townsville, Queensland, 4811, Australia. Electronic address: Leigh.owens@jcu.edu.au.
authors.forename: Leigh
authors.initials: L
authors.lastname: Owens
mesh_terms: D000818:Animals; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D017990:Densovirus / Q000235:genetics* / Q000302:isolation & purification; D033561:Penaeidae / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D045522:Recombinases / Q000378:metabolism*; D015203:Reproducibility of Results
chemical_list: D004279:DNA, Viral; D045522:Recombinases
keywords: Penaeus stylirostris; PstDV; PstDV-related sequences; RPA