狂犬病病毒抗体间接ELISA方法的初步建立与应用

Development and Preliminary Application of Indirect ELISA for Detection of Rabies Virus Antibodies

为建立一种提高犬狂犬病抗体检测水平的新方法,本研究选取狂犬病病毒G蛋白,将其信号肽删除,密码子优化后原核表达,得到重组狂犬病病毒糖蛋白G,然后将其作为包被抗原,建立检测犬血清抗体效价的间接ELISA方法;最后,用本试验建立的方法检测了92份临床样本,结果表明,犬的抗体水平与其免疫背景高度匹配,将部分样本与商品化试剂盒比对,一致性达90.9%,因此,该方法可用于检测犬血清中狂犬病病毒抗体水平,为疫苗免疫提供临床建议。

To develop a new method to improve the detection level of dog rabies antibodies,firstly,the signal peptide of virus G protein was deleted and its codon was optimized and then the recombinant glycoprotein G was expressed in prokaryotic expression system.The purified recombinant protein was used as the coating antigen and an indirect ELISA method was then developed for detecting the Rabies virus antibodies in dog serum samples.A total of 92 clinical dog serum samples was tested using this indirect ELISA method and a commercial ELISA kit.Both methods had 90.9%agreement on the test results.Based on the above results,the indirect ELISA method developed in this study might be used for clinical detection of rabies antibodies and providing clinical suggestions for vaccine immunity.