【摘要】 根据经典和新型水禽呼肠孤病毒σA基因的保守序列设计一对PCR引物,通过优化病毒RNA提取方法和PCR条件,建立了番鸭呼肠孤病毒通用RT-PCR检测方法。该方法可自1株经典型或4株新型水禽呼肠孤病毒特异性扩增出436 bp条带,其最低病毒检出量为3.6 TCID50,而对鸭甲肝病毒、鸭瘟病毒、新城疫病毒、坦布苏病毒、番鸭细小病毒、产蛋下降综合征病毒和H9N2亚型禽流感病毒扩增均为阴性。对2011~2012采集的224份(番鸭74、鹅68、鸭82)疑似临床病料进行检测,结果 56份为阳性,番鸭、鸭和鹅阳性率分别为59.5%(44/74),11.0%(9/82)和4.4%(3/68),表明水禽呼肠孤病毒主要在番鸭中流行。更多还原

【Abstract】 According to the conservative gene sequences of classical and novel waterfowl reovirus sigma A submitted in Gen Bank,a pair of universal primers were designed to carry out one-step RT-PCR amplification for detecting Muscovy duck reovirus. The PCR reaction condition was optimized and the ultimate one-step RT-PCR method had been established. A 436-bp PCR product was specifically amplified from 1 classical and 4 new waterfowl-origin reovirus strains with high sensitivity of 3. 6 TCID50 viruses,however,no product could obtain from duck hepatitis A virus,duck enteritis virus,Newcastle disease virus,Tembusu virus,Muscovy duck parvovirus,egg drop syndrome virus and H9N2 subtype avian influenza virus. With the established method,out of 224 clinical specimens from sick flocks of birds during 2011- 2012,a total of 56 were positive for reovirus. The positive incidences of Muscovy ducklings,ducklings and goslings were 59. 5%( 44 /74),11%( 9 /82) and 4. 4%( 3 /68),respectively. The results demonstrated that reovirus infection were mainly epidemic in Muscovy duck.更多还原