【摘要】 用2株不同致病力的H5亚型禽流感病毒(AIV)感染Madin-Darby canine kidney(MDCK)细胞,收集感染后6,12,24,48,72 h的病毒,提取总RNA,利用Rotor Gene RG3 000实时定量PCR仪对H5亚型AIV的HA基因进行检测,试图建立快速检测H5亚型AIV的real-time RT-PCR方法,并用所建立的方法对来自全国各地的疑似H5亚型AIV的病料和咽喉拭子、泄殖腔拭子346份进行检测。结果表明：农业部动物流感重点开放实验室建立的HA基因的teal-time RT-PCR方法可以检测到细胞培养物中和临床样品中是否含有H5亚型AIV,与常规RT-PCR比较,该方法更加特异、准确、快速。更多还原

【Abstract】 Two strains different pathogenicity H5 subtype AIV infected MDCK cells,total RNA were extracted from fluids after infected 6 h,12 h,24 h,48 h, 72 h and throat,tracheal swabs samples from chicken experimentally infected by H5 subtypes AIV,real-time RT-PCR of HA gene was done with Rotor Gene RG3000 PCR amplifier.We attempted to establish a real-time RT-PCR method to detect H5 subtype AIV.The results showed that RT-PCR method we eatablished could detect H5 subtype AIV of cell culture superoatant and swabs samples from chicken experimentally intfected.As such,the desoribed method is reliable and provides faster disease diagnostics than the conventional RT-PCR procedure to detect AIV.更多还原