Development of fluorescent real-time PCR method for detection of ASFV.

A specific primer pair and a probe were designed according to vp73gene from ASFV with an amplicon of 69 bp,and the fluorescent real-time PCR method in detecting ASFV was founded.Compared with three OIE reference methods,the results showed that four methods showed similar specificity.But the developed method was as sensitive as OIE real-time PCR method,but at least 10times more sensitive than the other two normal PCR methods.A quantitative assessment of standard curve showed the LLD of the developed method was at least 10copies/μL of virus ge- nome equivalents(R=0.993 046).Intra-assay and inter-assay CVvalues were 0.61%-1.14%and 1.37%-3.14% separately,which showed the method had good repeatability.With its high specificity,sensitivity and good repeat-ability,the developed method would become a valuable tool for the routine diagnostic procedures for ASFV.