【摘要】 依据新城疫病毒(NDV)融合蛋白(F)基因裂解位点的核苷酸序列与其毒力的相关规律,分别设计合成了四条寡核苷酸引物,建立了一个可迅速检测不同禽源新城疫毒株并可鉴定强、弱毒株的逆转录酶—聚合酶链式反应(RT—PCR)技术。对强毒株可扩增出362bp的通用片段和254bp的强毒株特异性片段;弱毒株则可扩增出362bp的通用片段和123bp的弱毒株特异性片段。本方法只需一个RT—PCR反应,整个过程在8小时内完成,既可检测感染鸡胚尿囊液,也可从病料直接检测。应用该方法对源于各种禽类的36份ND可疑病料样品进行检测试验,结果阳性检出率为73.5%,与常规方法检测的结果相一致。更多还原

【Abstract】 A reverse-transcriptase polymerase chain reaction(RT-PCR)based technique was developed to detect Newcastle disease virus(NDV)of different poultry species origin.Four oligonucleotide primers,based on the difference of nucleotide sequence at the cleavage site between virulent and non-virulent strains of NDV,were designed to amplify specific DNA from different virus.The result showed that samples confined non-virulent NDV can yield two bands of123bp and362bp,while those of virulent NDV can yield two bands of254bp and362bp.The RNA genome of virus used for RT-PCR can be isolated not only from the infected chicken embryonated,but also directly from tissue ho-mogenate of clinic samples.The detection results of36clinic samples of ND suspect were identical with the convention-al diagnosis in the study.The results demonstrated that the developed protocol of RT-PCR based is a specific?sensi-tive?rapid diagnostic technique for NDV of different poultry species origin and can also be used to differentiate the viru-lent and non-virulent strain.更多还原