| 摘要: |
| 【目的】利用MDBK细胞进行牛传染性鼻气管炎病毒(IBRV) XA株的分离与鉴定。【方法】用MDBK细胞从陕西发病奶牛鼻拭子中分离IBRV；使用IBR 标准阳性血清对该病毒分离物进行中和试验；根据GenBank公布的IBRV基因组序列，利用gB保守序列设计1对引物，对分离病毒进行PCR扩增并测序。【结果】分离病毒在MDBK细胞上产生了明显的特征性细胞病变（CPE）：细胞圆缩，聚集成葡萄串样群落，在单层细胞上形成空洞，偶见含有几个细胞核的巨大细胞；用标准抗IBRV特异性抗血清能完全中和分离株；用IBRV gB 特异性引物进行PCR，可扩增出1 182 bp的特异性条带，目的序列与已发表的IBRV基因组序列一致。【结论】该分离株为牛传染性鼻气管炎病毒，将其命名为 IBRV XA株。 |
| 关键词: BRV 分离鉴定 中和试验 PCR |
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| 基金项目:陕西省“13115”科技创新工程重大科技专项（2008ZDKG-05） |
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| Isolation and identification of infectious bovine rhinotracheitis virus |
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| Abstract: |
| 【Objective】The study was to isolate and identify infectious bovine rhinotracheitis(IBR) virus with Madin Darby bovine kidney (MDBK) cells.【Method】An infectious bovine rhinotracheitis(IBR) virus was isolated from a sick cow nasal swabs with (MDCK) cells,and the neutralizing antibody against the isolated virus was detected by the standard IBRV specific serum.A specific pair of primers were designed according to the gene sequence of IBRV gB gene publicated in GenBank.The gB gene of the IBRV strain was amplified by PCR method,and the sequences was analyzed．【Result】Obvious characteristics of the cytopathic effect(CPE) generated in MDCK,such as grape like clusters of rounded cells gathered around a hole in the monolayer;sometimes giant cells with several nuclei may be observed;The virus can be neutralized completely by the standard IBRV specific serum.Specific band can be amplicified by PCR with IBRV gB specific primers and the sequencing result showed that the sequence of the isolated virus coincided with the published IBRV gB sequences.【Conclusion】The result indicated that the isolate virus was IBRV,and was named IBRV XA. |
| Key words: Infectious bovine rhinotracheitis virus(IBRV) isolation and identification neutralization test PCR |
