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
杨敩校 1刘芊麟 1陈丙生 1蒋小玲 2王建舫 1董虹 1肖兴 2周双海1
作者信息
- 1. 北京农学院动物科学技术学院,北京 102206
- 2. 衡阳市畜牧水产事务中心,湖南衡阳 421001
- 折叠
摘要
[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.
Abstract
[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 in-tra-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.
关键词
猪细环病毒1型/衣壳蛋白/间接ELISA/抗体/检测Key words
TTSuV1/capsid protein/indirect ELISA/antibody/detection引用本文复制引用
基金项目
国家自然科学基金资助项目(31340050)
北京市家畜创新团队项目(BAIC05-2023)
出版年
2024
NETL
NSTL
万方数据