Development and application of a TaqMan real-time fluorescence quantitative PCR for serotype 1 FAdV
To detect serotype 1 FAdV infection in chickens,primers and TaqMan probes were de-signed according to the conserved 52K gene of serotype 1 FAdV in this study.Real-time fluores-cence quantitative PCR optimized by the matrix method was used to detect the virus shedding of o-ral and cloacal swabs and the viral load of tissues after challenge.The results showed that the method had good specificity,sensitivity,and reproducibility;it could only detect serotype 1 FAdV,but not other serotypes of FAdV or other avian viruses.The minimum detection limit was as low as 10 copies/μL,and the inter-and intra-group dispersions were less than 2％.The standard curve was y=-3.836x+41.791,and the correlation coefficient was R2=0.996,indicating a good linear relationship.The results of the SPF chicken challenge showed that FAdV serotype 1 could be de-tected in the oral and cloacal swabs until 14 days post-infection,and the virus could also be detec-ted in the liver,kidney,gizzard,and ileum at 8 days post-infection,but not in the glandular stomach and cecum.This study provides a method for the quantitative detection of serotype 1 FAdV.
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