生物学杂志

生物学杂志

• 技术方法 • 上一篇    下一篇

O型口蹄疫病毒抗体高通量液相阻断CLIA检测方法的建立

  

  1. 1. 江南大学 粮食发酵工艺与技术国家工程实验室， 无锡 214122；2. 无锡市人民医院, 无锡 214000； 3. 中农威特生物科技股份有限公司， 兰州 730046
  • 出版日期:2018-12-18 发布日期:2018-12-18
  • 通讯作者: 白仲虎，教授，主要从事体外诊断研究工作，E-mail：baizhonghu@jiangnan.edu.cn
  • 作者简介:王世杰，硕士研究生，从事体外诊断试剂研究，E-mail：594149292@qq.com； 王春新，副教授，从事临床检验诊断学研究，E-mail：wangcx_wxph@163.com
  • 基金资助:
    863课题（2015AA020802）

Establishment of high throughput liquid-phase blocking CLIA for detecting antibodies to foot-and-mouth disease virus type O#br#

  1. 1. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122;2. Wuxi People′s Hospital, Wuxi 214000; 3. China Agricultural Vet. Bio. Science and Technology Co., Ltd., Lanzhou 730046, China
  • Online:2018-12-18 Published:2018-12-18

PDF (PC)

135

摘要： 为了能更高效和准确地对口蹄疫疫情进行诊断监测，建立了O型口蹄疫病毒抗体高通量液相阻断化学发光免疫定性检测方法（Liquid-phase blocking CLIA，LPB-CLIA）。通过以单一稀释倍数稀释样本进行检测，计算抑制率作为检测结果，将检测通量提高了5倍左右。应用受试者工作特征曲线（Receiver operating characteristic，ROC）确定试验最佳临界值为62.5%。重复性实验结果显示方法批内CV小于10%，批间CV小于15%。通过对326份田间血清样本检测结果与LPB-ELISA检测结果对比，敏感性为94.6%，特异性为95.6%，检测准确度为95.1%。建立的LPB-CLIA方法敏感性和特异性良好，缩减了时间和经济成本，为国内口蹄疫的检疫提供了新的方法。

关键词: 口蹄疫, 液相阻断, 化学发光, 高通量, 抗体检测

Abstract: In order to diagnose and monitor FMD(foot-and-mouth disease) efficiently and accurately, this study established a high throughput liquid-phase blocking CLIA antibody detection method of the O-type FMD. Through using one single dilution instead of the gradient dilution, the detection flux was raised about 5 times. The test results were assessed by percentage inhibition, and the optimal cut-off value was 62.5% determined by the receiver operating characteristic curve (ROC). Repeatability test showed the intro batch CV was under 10% and the inter batch CV was under 15%. By comparing the results of 326 field serum samples with those of LPB-ELISA, the sensitivity, sepecificity and accuraly were 94.6%, 95.6% and 95.1%, respectively. The results showed that the kit reduce the time and economic cost on the basis of the good sensitivity, specificity and reliability of the results. It also provides a new method for the quarantine of foot and mouth disease in China.

Key words: FMD, liquid-phase blocking, CLIA, high throughput, antibody detection

中图分类号: