【摘要】 为实现牛病毒性腹泻病毒(BVDV)与牛肠道病毒(BEV)的同步检测，靶向BVDV和BEV 5′-UTR序列保守区域，分别设计1对引物，建立针对BVDV与BEV的SYBR GreenⅠ双重qPCR检测方法。结果显示，该方法可特异性检出BVDV和BEV,两者熔解峰所对应的Tm值分别为87、82℃,而对其他常见牛病原体无特定熔解峰。BVDV和BEV的最低检测值分别为81.3、81.7拷贝/μL;批内及批间的相对标准偏差均被有效控制在2%的限定水平。运用建立的SYBR GreenⅠ双重qPCR检测方法对2023—2024年采自河南、山东以及四川等地牛场的44份粪便样品进行检测，BVDV和BEV的检出率分别为75.00%(33/44)和77.27%(34/44),两者共感染检出率为50%(22/44)。该方法为BVDV和BEV同步检测提供了一种快速、灵敏且特异的解决方案。更多还原

【Abstract】 In order to detect simultaneously bovine viral diarrhea virus(BVDV) and bovine enterovirus(BEV),two pairs of specific primers were designed based on the conserved regions of the 5’-UTR sequences of BVDV and BEV.A SYBR Green Ⅰ-based duplex qPCR assay for simultaneously detecting BVDV and BEV was established.The results showed that BVDV and BEV could be specifically detected by this assay,with distinct melting peaks of 87 ℃ for BVDV and 82 ℃ for BEV,respectively,while no specific melting peaks were exhibited for other common bovine pathogens.The lowest detection values of BVDV and BEV were determined to be 81.3 copies/μL and81.7 copies/μL,respectively.Both intra-and inter-assay coefficients of variation were maintained below 2%.The SYBR Green Ⅰ-based duplex qPCR assay was then applied to detect 44 fecal samples collected from cattle farms in Henan,Shandong and Sichuan Provinces during 2023—2024,and the detection rates of BVDV and BEV were 75.00%(33/44) and 77.27%(34/44) respectively,whereas the detection rate of co-infection for BVDV and BEV was 50%(22/44).This optimized procedure establishes a rapid,sensitive,and highly specific detection method for the simultaneous identification of BVDV and BEV.更多还原