猪细环病毒1型间接ELISA抗体检测方法的建立

Development of an indirect ELISA for detection of antibody to Torque teno sus virus type 1

【目的】建立一种检测猪细环病毒1型抗体的血清学方法。【方法】以猪细环病毒1型衣壳蛋白为包被抗原,筛选和优化抗体检测反应条件来建立一种检测猪细环病毒1型衣壳蛋白抗体的间接ELISA方法,之后进行特异性和重复性试验,并进行初步临床应用检测。【结果】确定了检测猪细环病毒1型衣壳蛋白抗体的间接ELISA方法的各项最佳反应条件;对8种猪源病毒阳性血清的检测结果显示只有猪细环病毒1型阳性血清为阳性,显示出良好的特异性;批内变异系数均小于5%,批间变异系数均小于10%,显示出良好的重复性。对来源于北京和湖南地区的400份猪血清样品的检测结果显示,猪细环病毒1型衣壳蛋白抗体阳性率为75.25%,PCR阳性率为71.00%,二者之间没有显著差异(P>0.05),表明这两个地区存在较多的的猪细环病毒1型感染。【结论】建立了一种特异性和重复性都良好的检测猪细环病毒1型衣壳蛋白抗体的间接ELISA,可用于猪细环病毒1型抗体的临床检测。

[Objective]This study was aimed to establish a serological method for detection of Torque teno sus virus type 1(TTSuV1).[Methods]Using the capsid protein of TTSuV1 as coating antigen,the reaction conditions for antibody detection were screened and optimized to establish an indirect enzyme-linked immunosorbent assay(ELISA)for detection of the antibody against TTSuV1 capsid protein.Subsequently,its specificity and reproducibility test were conducted,and its preliminary clinical detection were performed.[Results]All the best reaction conditions of the indirect ELISA for the detection of the antibody to TTSuV1 capsid protein were determined.The detection results of 8 kinds of positive serum samples of swine-derived viruses showed that only positive serum samples of TTSuV1 was positive,showing good specificity.The coefficients of variation for intra-assay was under 5%,and that for inter-assay was under 10%,indicating good reproducibility.The detection results of 400 pig serum samples from Beijing and Hunan regions showed that the positive rate of antibodies against TTSuV1 capsid protein was 75.25%,and the positive rate of PCR was 71.00%,there is no significant(P>0.05)difference between the two methods,and which indicated that there was a high rate of TTSuV1 infection in this two regions.[Conclusion]An indirect ELISA with good specificity and reproducibility for detecting antibodies to the TTSuV1 capsid protein was established,and it can be used for clinical detection of antibodies against TTSuV1.