蚀斑法分离纯化BHV-1和BPIV-3混合病毒

发布时间：2018-05-22 12:29

本文选题：牛疱疹病毒型 + 牛副流感病毒型　；参考：《中国兽医杂志》2017年01期

【摘要】：利用蚀斑试验方法对牛疱疹病毒1型(BHV-1)和牛副流感病毒3型(BPIV-3)的混合病毒进行分离纯化及鉴定,为BHV-1、BPIV-3感染的诊断提供科学依据。将不同稀释度的BHV-1和BPIV-3混合病毒接种至牛肾原代细胞,加营养琼脂培养,产生蚀斑后扩增并RT-PCR鉴定。将纯化后毒液分别进行二次蚀斑纯化及鉴定。结果 BHV-1和BPIV-3混合病毒第6~7 d可产生明显蚀斑。鉴定阳性的BHV-1和BPIV-3进行二次蚀斑,鉴定结果均为阳性,且分离的两种病毒TCID_(50)测定结果为10~(-8.5)/0.1 m L、10~(-6.5)/0.1 m L。首次通过蚀斑试验,成功分离并得到了两株毒力较高且纯化的BHV-1和BPIV-3病毒株。
[Abstract]:The mixed viruses of bovine herpesvirus 1 (BHV-1) and bovine parainfluenza virus type 3 (BPIV-3) were isolated and identified by plaque test, which provided scientific basis for the diagnosis of BHV-1 and BPIV-3 infection. The mixed virus of BHV-1 and BPIV-3 with different dilution was inoculated into bovine kidney primary cells and cultured with nutritional Agar. The plaque was produced and then amplified and identified by RT-PCR. The purified venom was purified and identified by secondary plaque purification. Results BHV-1 and BPIV-3 mixed virus could produce obvious plaque on the 6th day. The positive BHV-1 and BPIV-3 were identified as secondary plaque, and the results were both positive. The results of the two isolated viruses TCID-50) were 10 ~ 8.5 / 0.1 mL / 10 ~ (-6.5) / 0.1 ~ (-1) ml / 0 ~ (-1) 路m ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) ~ (-1) ~ (-1) ~ (-1). Two highly virulent and purified BHV-1 and BPIV-3 viruses were successfully isolated and purified by plaque test for the first time.
【作者单位】：西北民族大学生命科学与工程学院;生物工程与技术国家民委重点实验室;
【基金】：教育部“长江学者和创新团队发展计划”项目(IRT13091) 校企合作项目(兰州民海生物工程有限公司,XBMU-2014-BC-18) 中央专项研究生科研创新项目(Yxm2015210)
【分类号】：S852.65

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