Development and Application of TaqMan Real-Time Quantitative PCR for Detection of African Swine Fever Virus K205R Gene
In order to rapidly and accurately detect African swine fever virus(ASFV),this study designed and synthesized primers and probes based on the published K205R gene sequence of the prevalent strain ASFV SY18(GenBank ID:MH766894.1)in China and developed a TaqMan fluorescent quantitative PCR assay.The Ct value of the assay showed a good linear relationship with the standard in the range of 1×1011 to 1×102 copies/μL with a correlation coefficient of 0.998 and a slope of-2.797.The lower limit of detection was 102 copies/μL and there was no cross-reactivity with other porcine viruses that caused similar signs.In addition,the coefficients of variation between and within groups was less than 1.53774%.In summary,the TaqMan fluorescent quantitative PCR assay developed in this study had good specificity,sensitivity and reproducibility,which could be used for early diagnosis of ASF and rapid detection of ASFV and recombinant porcine reproductive and respiratory syndrome virus expressing ASFV K205R protein.
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