SHI Wei-Wen and DU Nian-Xin. Purification of Rabbit Haemorrhagie Disease Virusfrom Counter Immunoelectrephoresis[J]. Virologica Sinica, 1995, 10(3).
Citation: SHI Wei-Wen, DU Nian-Xin. Purification of Rabbit Haemorrhagie Disease Virus from Counter Immunoelectrephoresis .VIROLOGICA SINICA, 1995, 10(3) : 261.

用对流电泳提纯兔出血症病毒

  • 兔出血症病毒(RHDV)在pH8.6的琼脂糖凝胶上电泳,在负极侧有一血凝峰,免疫电镜观察有大硅RHDV颗粒,证实RHDV病毒粒子带正电荷。利用这一特性可用对流电泳提纯病毒。将抗体与病毒颗粒形成的沉淀带切下,作SDS-PAGE,经免疫转印出现6条区带,其中60kD的VP_1为主要结构多肽。用免疫复合物提取核酸,以狄高辛标记制成探针,能与病毒核酸和克隆的RHDVcDNA2.2kb片段和4.8kb片段杂交,探针灵敏度达pg水平。能用病毒核酸作模板制备探针,证实RHDV的核酸为DNA。

Purification of Rabbit Haemorrhagie Disease Virus from Counter Immunoelectrephoresis

  • HDV could be moved to the negative electrode by electrophoresis in the condition ofpH8.6 ,and this was confirmed by immunoelectron microscopy observation. The RHDV im-munocomplex was purified by counter immunoelectrophoresis(CIE)form liver tissue of rab-bits died of RHD and used for the analysis of RHDV structural polypeptide by immunoblot-ing.The RHDV nucleic acid was extracted from RHDV immunocomplex and labelled withDigoxigenin-dUTP. The labelled nucleic acid probe with high senseitivity and specificity wasable to hybrideized with RHDV nucleic acid,RHDV 2. 2kb and4. 8kb cDNA fragments in dotblot analysis.

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    Purification of Rabbit Haemorrhagie Disease Virus from Counter Immunoelectrephoresis

    • 1. Medical College of Animals, Nanjing Agricultural University

    Abstract: HDV could be moved to the negative electrode by electrophoresis in the condition ofpH8.6 ,and this was confirmed by immunoelectron microscopy observation. The RHDV im-munocomplex was purified by counter immunoelectrophoresis(CIE)form liver tissue of rab-bits died of RHD and used for the analysis of RHDV structural polypeptide by immunoblot-ing.The RHDV nucleic acid was extracted from RHDV immunocomplex and labelled withDigoxigenin-dUTP. The labelled nucleic acid probe with high senseitivity and specificity wasable to hybrideized with RHDV nucleic acid,RHDV 2. 2kb and4. 8kb cDNA fragments in dotblot analysis.

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