Establishing a gold standard method for the detection of Cherax reovirus using reverse transcriptase, quantitative, polymerase chain reaction

Cherax reovirus infects redclaw crayfish (Cherax quadricarinatus) and it may be involved in mortalities between 5-20 % stunting of up to 40 survivors. The sequence the RNA-dependent RNA polymerase was used develop a reverse transcription, quantitative, PCR (RT-qPCR) which specific against seven other crustacean viruses (Athtab bunyavirus, Chequa iflavirus, Macrobrachium rosenbergii nodavirus, Gill-associated virus, Taura syndrome White spot Penaeus stylirostris Penstylhamaparvovirus) although GAV produced reaction that easily separated by melt curve analysis. A strong linear correlation (r2 = 0.9965) obtained viral quantities ranging from 107 10 copies/reaction with an amplification efficiency 0.92. This RT-qPCR is 2-times faster 100 times more sensitive than standard RT-PCR using agarose gel electrophoresis potential detect virus down 7.64 clinical samples. In crayfish samples, able when traditional negative. Its' measurement uncertainty less 2% (0.02-1.9), similar PCRs for viruses. proposed as gold should screening populations C. quadricarinatus broodstock before being hatcheries or on farms.