中文摘要：

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。

英文摘要：

In order to provide scientific basis for BHV-1, BPIV-3 diagnosis of infection , we utilized the plaque test for mixed virus isolation, identification and purification of bovine herpesvirus type 1 （ BHV-1 ） and bovine parainfluenza virus type 3 （ BPIV-3 ）, The various dilutions of BHV-1, and BPIV-3 mixed viruses were inoculated into a dense monolayer primary calf kidney cell. and the cells were then covered with nutritious agar. After generating plaques, the most obvious CPE clones were selected to be identified by RT-PCR reaction. Purification and identification of the purified venom were performed for two times. The plaques of BHV-1 and BPIV-3 mixed viruses were revealed after 6 to 7 days. Positive identification of BHV-1 and BPIV-3 was performed second plaque, and the results were still positive. The TCID50 of the two virus isolates were 10 ^-8.5/0. 1mL and 10^ -6.5/0.1mL. For the first time, the two strains of BHV-1 and BPIV-3 with high virulence were successfully separated and purified by plaque assay.