鸭瘟病毒PCR检测方法的建立及验证

目的建立鸭瘟病毒PCR检测方法,并进行验证。方法根据GenBank中登录的鸭瘟病毒基因(UL30、UL31)序列,设计合成1对特异性引物,以鸡胚化弱毒疫苗株病毒基因组DNA为模板,进行PCR扩增。优化PCR的反应条件,并对方法的敏感性、特异性及适用性进行验证。结果经PCR可扩增得到446 bp的目的基因条带,测序结果与已发表的序列同源性达100%。该方法的最低检出限为2.1 pg,对7种鸭易感性病原体及3种鸭瘟病毒同类病原体的检测结果均为阴性。结论已建立了敏感、特异、快速的鸭瘟病毒PCR检测方法。

Development and Verification of PCR for Detection of Duck Plague Virus

Objective To develop and verify a PCR method for detection for duck plague virus.Methods A pair of specific primers were designed according to the sequence of duck plague virus genes UL30 and UL31 reported in GenBank for PCR using the viral genome of attenuated vaccine strain adapted in chick embryo as a template,based on which the reaction condition for PCR was optimized,and the sensitivity,specificity and adaptivity of the developed method were verified.Results The target gene band at a length of 446 bp was amplified by PCR,with a homology of 100% to the gene sequence reported in GenBank.The minimum detection limit of the developed method was 2.1 pg.All the detection results of 7 pathogens susceptible to duck and 3 congeneric pathogens of duck plague virus by the developed method were negative.Conclusion A sensitive,specific and rapid PCR method for detection of duck plague virus was developed.