根据鸭甲肝病毒(Duck hepatitis A virus,DHAV)的3D基因序列和新型鸭呼肠孤病毒(Novel Duck reovirus,NDRV)的S3基因序列,分别设计了1对特异性引物,通过条件优化建立了鉴别DHAV和NDRV的复合RT-PCR方法。该方法对DHAV和NDRV的最低检出量均为100 copies,而对番鸭呼肠孤病毒(Muscovy duck reovirus,MDRV)、鸭瘟病毒(Duck plague virus,DPV)、鸭坦布苏病毒(Duck Tembusu virus,DTMUV)和鸭副粘病毒(Duck paramyxo virus,NDV)的检测结果均为阴性。临床样品的检测结果显示,该方法是一种快速鉴别诊断DHAV和NDRV感染的有效手段。
A multiplex reverse transcription polymerase chain reaction(RT-PCR) assay for detecting Duck hepatitis A virus(DHAV) and Novel duck reovirus(NDRV) was developed using two primers based on 3D gene of DHAV and S3 gene of NDRV. The detection limit of the RT-PCR assay was 100 copies for both viruses. Mean while, the method had no reaction with Muscovy duck reovirus(MDRV), Duck plague virus(DPV), Duck Tembusu virus(DTMUV) and Duck paramyxo virus(NDV). The results of testing clinical samples showed the assay was a useful technique for differential detection of DHAV and NDRV.