Abstract: In order to develop a credible dual fluorescence quantitative polymerase chain reaction（qPCR）for detecting novel goose astrovirus type 1（NGAstV⁃1）and type 2（NGAstV⁃2），primers were designed to establish a dual fluorescence qPCR detection method using ORF2 protein of NGAstV⁃1 and NGAstV⁃2，and 36 samples of goose liver and kidney tissue suspected to be infected with NGAstV⁃1 and NGAstV⁃2 were evaluated in this study. The test results showed that the correlation coefficients of qPCR were 0.997 and were negative for novel duck reovirus （NDRV），avian influenza virus（AIV），fowl adenovirus（FAdV），and goose parvovirus（GPV）. The minimum detection limit was 1.12 × 101 copies/μL. The reproducibility test results indicate good reproducibility，with a coefficient of variation between groups of less than 0.8% and a coefficient of variation within groups of less than 0.5%. It can be seen that the qPCR can efficiently and specifically detect NGAstV⁃1 and NGAstV⁃2，providing a feasible method for NGAstV monitoring.

Key words: Novel goose astrovirus, qPCR, ORF2

摘要： 为建立一种行之有效地检测新型鹅星状病毒（Novel Goose Astrovirus，NGAstV）基因 1型（NGAstV-1）和基因2型（NGAstV-2）双重荧光定量PCR（quantitative polymerase chain reaction，qPCR），该研究根据NGAstV-1 和NGAstV-2 的ORF2 蛋白设计鉴定引物，建立SYBR Green I的双重qPCR检测方法并用36份疑似感染NGAstV-1和NGAstV-2的雏鹅肝及肾组织样 品进行评估。试验结果显示qPCR相关系数为0.997，对鸭坦布苏病毒（duck tembusu virus，DT- MUV）、番鸭呼肠孤病毒（muscovy duck reovirus，MDRV）、新型鸭呼肠孤病毒（novel duck reovi? rus，NDRV）、禽流感病毒（avian influenza virus，AIV）、禽腺病毒（fowl adenovirus，FAdV）、鹅细小 病毒（goose parvovirus，GPV）扩增结果均为阴性；最低检测限度为1.12×101 copies/μL；重复性试 验结果显示：组间变异系数小于0.8%，组内变异系数小于0.5%，具有良好重复性。由此可知， 该qPCR可高效、特异地检测NGAstV?1和NGAstV?2，为NGAstV监控提供切实可行的方法。

关键词: 新型鹅星状病毒, qPCR, ORF2