牛病毒性腹泻—黏膜病病毒分离鉴定及生物学特性研究

doi:10.16431/j.cnki.1671-7236.2015.09.027

Abstract

Abstract: In order to isolate and identify bovine viral diarrhea-mucosal virus (BVDV) strain of Xinjiang, grasp the biological characteristics of the strain, we gathered feces which were suspected as bovine viral diarrhea-mucosal disease infected feces in the northern parts of Xinjiang.The strain was separated and identified by RT-PCR, cell isolation and culture, indirect immunofluorescence antibody test, immune electron microscopy and serum neutralization test.After determining TCID₅₀ of the strain, ether sensitivity test, chloroform sensitivity test, trypsin sensitivity test, acid and alkali sensitivity test, temperature sensitivity test and nucleic acid typing test were used to detect the physical and chemical properties of the strain.The samples appeared fragment at 286 bp by RT-PCR, and then inoculated the positive samples into a density of about 80% monolayer MDBK cells, and appeared cytopathic effect, passaged to the 5th generation caused the typical cytopathic effect.The F5 cell cultures were detected by indirect immunofluorescent antibody, the result showed that a yellow-green fluorescence appeared same as C₂₄V.A lot of spherical BVDV particles with the size of 20 to 40 nm were observed by immune electron microscopy.Serum neutralization test had no cytopathic effect, viruses were completely neutralized by positive antibody serum.The isolated strain was determined to be BVDV based on the methods above.The TCID₅₀ of the isolated strain was 10^-4.5/0.1 mL.Physicochemical assays showed that the isolated strain was sensitive to chloroform, ether, trypsin and temperature, sensitive to acid and resistant to alkali in acid and alkali sensitivity test, and was completely inactivated by 54 ℃ for 1 h.The virus genome was RNA.The study successfully isolated a BVDV strain in Xinjiang and grasped the biological characteristics of the strain, which laid a foundation for future diagnosis and prevention of the disease.

Key words: bovine; bovine viral diarrhea-mucosal disease virus (BVDV); isolation and identification; biological characteristics

CLC Number:

S858.23

BAO Zhen-zhong, LEI Cheng-hong, CAI Yuan-qing, GAO Dou, BIAN Sai-sai, GE Ting. Isolation,Identification and Biological Characteristics Analysis of Bovine Viral Diarrhoea-mucosal Disease Virus[J]. , 2015, 42(9): 2391-2398.

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Isolation,Identification and Biological Characteristics Analysis of Bovine Viral Diarrhoea-mucosal Disease Virus

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