鹅细小病毒与番鸭细小病毒PCR鉴别诊断方法的建立

Establishment of PCR Assay for Differentiation of Goose Parvovirus from Muscovy Duck Parvovirus

根据GenBank上登录的鹅细小病毒(GPV)和番鸭细小病毒(MDPV)的基因序列,针对两种病毒非结构蛋白(NS)和VP1基因序列的相同区段,分别设计两对引物GPV U.L-1和MDPV U.L-1,以GPV-GZ1株的鹅胚尿囊液和MDPV的番鸭胚尿囊液提取核酸作为模板,分别建立了检测GPV和MDPV的PCR方法。特异性试验结果显示,引物GPV U.L-1仅特异性扩增出GPV-GZ1和GPV-GZ2株分子长为622 bp核酸片段,引物MDPVU.L-1仅特异性扩增出MDPV分子长为624 bp核酸片段,而对DPV、GPMV的核酸扩增结果均为阴性;敏感性试验结果显示,建立的PCR方法能检测到0.144 pg的GPV核酸和28.8 pg的MDPV核酸。结果表明,建立的PCR方法特异性强、敏感性高,可用于GPV或MDPV临床感染病例的鉴别诊断。

Two pairs of primers （U·L-1） of goose parvovirus （GPV） and Muscovy duck parvovirus （MDPV） were designed based on the conserved NS/VPlsequence of GPV and MDPV released in website, respectively. Nucleic acids of GPV - GZ1 and MDPV strains were extracted respectively from aUantoic fluids of goose and muscovy duck embryo. Sub- sequenfly, PCR assay were developed successfully for detection of the two kinds of virus. The specific fragments of 622 bp and 624 bp in size were amplified from DNA samples of GPV - GZ1 or GPV - GZ2 and MDPV, respectively, rather than that of DPV and GPMV. DNA sensitivity used for PCR detection might be as low as 0. 144 pg for GPV or 28. 8 pg for MDPV DNA. The results showed that the PCR technique was rapid, sensitive and specific for the differential diagnosis of GPV and MDPV infections. Key words： Goose parvovirus （GPV）, Muscovy duck parvovirus （MDPV）, PCR, Differential diagnosis