Development of a TaqMan® PCR assay with internal amplification control for the detection of African swine fever virus

摘要: Abstract A closed-tube polymerase chain reaction (PCR) was developed to allow the rapid detection of African swine fever virus (ASFV) DNA. This assay targets VP72 gene ASFV and uses 5′-nuclease (TaqMan®) system detect PCR amplicons, avoiding tube opening potential cross-contamination post-PCR products. An artificial mimic engineered with TaqMan® probe site replaced by a larger irrelevant DNA fragment allowing discrimination from using two-colour reporters. When added samples, successful amplification this demonstrated absence substances inhibitory PCR, thereby validating negative results. Assay sensitivity confirmed obtaining positive signals representative selection isolates. Many clinical post-mortem features ASF resemble those classical (CSF) porcine dermatitis nephropathy syndrome (PDNS). Therefore, fast reliable is essential not only for implementation control measures prevent spread ASF, but also in differential diagnosis CSF PDNS. should prove be valuable tool laboratory will complement existing molecular methods provide cases suspected fever.