脊髓灰质炎病毒I型抗体胶体金检测试纸条的研制
Development of poliovirus type I antibody colloidal gold test strip
投稿时间:2019-05-05  修订日期:2019-05-05
DOI:
中文关键词: 脊髓灰质炎病毒  单克隆抗体  胶体金试纸条
英文关键词: poliovirus  monoclonal antibody  colloidal gold test strip
基金项目:中央高校基本科研业务费专项资金资助项目(37001/CZP17010);中南民族大学横向科研资助项目(HZY17059)
作者单位E-mail
王朝元 中南民族大学生命科学院 wangchaoyuan@mail.scuec.edu.cn 
黄圣 中南民族大学生命科学院  
尹新涛 武汉生命科技股份有限公司  
项雅利 武汉生命科技股份有限公司  
尹玉莹 中南民族大学生命科学院  
莫娇 中南民族大学生命科学院  
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中文摘要:
      通过胶体金免疫层析技术建立一种特异、快速的脊髓灰质炎病毒抗体检测方法,对接种脊灰疫苗后的抗体水平监测提供参考.用辛酸-硫酸铵沉淀法纯化脊灰单抗,用柠檬酸三钠还原法制备的胶体金,标记纯化单抗,再结合脊灰抗原形成复合物,固定于金标垫上,将鼠抗人IgG和羊抗鼠IgG分别包被于试纸条的 T (检测线) 处和 C (质控线) 处,组装试纸条.应用该试纸条进行临床样品检测.结果表明,以稀释后的脊灰阳性血清作为检测样品,试纸条检测的灵敏度为1:80;特异性试验证明该试纸条与甲型肝炎病毒、乙型肝炎病毒、麻疹病毒抗体阳性血清没有交叉反应;在4℃和室温条件下保存3个月,试纸条的灵敏度和特异性没有明显改变,说明试纸条稳定性良好;检测结果与脊灰疫苗抗体检测试剂盒(ELISA)试验的符合率为86.7%.表明该脊灰病毒抗体胶体金检测试纸可以方便、快速、灵敏、特异性地检测脊灰病毒的抗体水平,适用于临床大规模血清抗体水平监测,具有良好的应用前景.
英文摘要:
      A specific and rapid detection method of poliovirus antibody was established by colloidal gold immunochromatography, which provides a reference for monitoring antibody levels after polio vaccination. Polio monoclonal antibody was purified with caprylic acid-ammonium sulfate precipitation method and colloidal gold was prepared by trisodium citrate reduction method. Colloidal gold labeled purified monoclonal antibody was combined with polio antigen to form complex, which immobilized on gold standard pad. Mouse anti-human IgG and the goat anti-mouse IgG was coated on the T (test line) and C (quality control line) of the test strip respectively, and the test strip was assembled. The test strip was used for sample test and the results showed that the sensitivity of the test strip was 1:800 after the diluted polio-positive serum was used as the test sample; the specificity test results proved that the test strip did not cross-react and the hepatitis A virus, Hepatitis B virus and measles virus antibody positive serum; after stored for 3 months at 4 ℃ and room temperature, the sensitivity and specificity of the test strip did not change significantly, indicating that the test strip was stable; The coincidence rate of our results with polio-vaccine antibody detection kit (ELISA) test was 86.7%, which confirmed that the established poliovirus antibody colloidal gold test strip is a conveniently, rapidly, sensitively and specifically method to detect the antibody level of poliovirus. It is suitable for clinical large-scale serum antibody level monitoring, and has good application prospect.
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