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自英国引进3株荧光素标记的针对狂犬病毒核蛋白的单克隆抗体,建立狂犬病直接免疫荧光诊断技术。根据已知狂犬病毒核蛋白(N或NP)基因序列,设计合成4对引物,建立狂犬病RT-PCR及套式PCR诊断技术。自牟定采集发病犬脑组织样品15份,免疫荧光及PCR诊断结果均为阳性。对PCR产物进行纯化后,克隆至pMD18-T载体测序(基因库登录号:EU095330),并与已知代表毒株对应序列进行比对及系统发育分析,结果表明:云南牟定狂犬病毒属于基因1型和血清1型毒株,与2006年广西发病犬分离毒株遗传关系密切,病毒部分N基因核苷酸及推导的氨基酸序列同源性分别为97.4%和99.0%。
Since the introduction of three fluorescein-labeled monoclonal antibodies to rabies virus nucleoprotein in the UK, rabies immunofluorescence was established. According to the known sequence of rabies virus nucleoprotein (N or NP), four pairs of primers were designed and synthesized, and Rabies RT-PCR and nested PCR were established. Self-Mouding collected dog samples of brain tissue samples 15, immunofluorescence and PCR diagnostic results were positive. The PCR product was purified and cloned into the pMD18-T vector (GenBank accession number: EU095330). The PCR products were aligned with the corresponding sequences of known representative strains and phylogenetically analyzed. The results showed that: 1 and serotype 1 strains were closely related to the isolates of dogs isolated from Guangxi in 2006. The homologies of the nucleotide and deduced amino acid sequences of the partial N gene were 97.4% and 99.0%, respectively.