稳定表达SLAM受体的Vero和BHK21细胞系的建立及其对犬瘟热病毒分离效果的比较

doi:10.11843/j.issn.0366-6964.2018.06.013

Abstract

Abstract:

The objective of this work was to establish a SLAM-Vero cell line and a SLAM-BHK21 cell line that stanbly expressing SLAM receptor, and then to compare their sensitivities to the canine distemper virus (CDV) wild strains, aiming at providing another tools for the rapid isolation of CDV for further study. The eukaryotic expression vector Pcag-SLAM, which contains the SLAM gene, was transfected into the Vero cells and BHK-21 cells, respectively. Then the cells which harbor the SLAM gene were subsequently screened with G418 combined with end point dilution, RT-PCR, immunofluorescence assay, and Western blot. The two cell lines were subsequently applied to isolate virus from 5 different tissues in 3 clinical cases. The isolated CDV was further identified by RT-PCR and Indirect Immunofluorescence assay. The results showed that the SLAM-Vero cells lines and SLAM-BHK21 cells lines were constructed successfully. SLAM-Vero inoculated virus for 12-24 h presented a typical CDV-induced cell fusion of CPE. By using SLAM-Vero cells, two strains of CDV were obtained from three CDV positive canine lungs and spleens, while SLAM-BHK21 cell lines, parental Vero cells and parental BHK21 cells failed to isolate the virus. The results indicated that CDV was more readily to replicate and proliferate in SLAM-Vero cells and easier to produce visible CPE than that in SLAM-BHK21 cells. Canine lungs and spleens are more ideal candidates for isolation of CDV when using the newly established SLAM-Vero cell lines in this report.

CLC Number:

S852.659.5

ZHU Lu, YAN Xia, DAI Ke, WANG Zheng-hao, ZHAO Yu-jia, YANG Zhen, WEN Xin-tian, CAO San-jie, HUANG Xiao-bo, WU Rui, ZHAO Qin, WEN Yi-ping. Establishment of Vero and BHK21 Cell Lines with a Stable Expression of Canine Distemper Virus Cell Receptor SLAM and Comparison of Their Effects on the Isolation of Canine Distemper Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(6): 1212-1221.

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Establishment of Vero and BHK21 Cell Lines with a Stable Expression of Canine Distemper Virus Cell Receptor SLAM and Comparison of Their Effects on the Isolation of Canine Distemper Virus

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