ObjectiveTo apply SYBR green I fluorescent quantitative real-time PCR (QPCR) in detecting 4 vertical transmission viruses of chicken and to evaluate its application value for the quality control of SPF chickens. MethodQuantitative real-time PCR technique was used to detect REV, ALV, ARV and CAV RNA, DNA or provirus DNA in 30 cloacal swab samples and 30 eggs of SPF chicken samples as well as 40 cloacal swab samples and 30 eggs of common chicken samples. Standard curve and melting curve analyses were conducted to obtain the quantitative result. ResultsOut of the SPF chicken samples, 2/30 (3.3%) were ALV-positive, and the three other viruses were negative. For the common chicken samples, 2/70 (2.9%) were REV-positive, and 10/70 (14.3%) were ALV-positive. ConclusionsThese QPCR assays show better sensitivity and specificity for detecting the 4 viruses in SPF chickens with a detection limit of 100 copies of target DNA per-reaction, and have a bright prospect in clinical application.