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结核、血吸虫抗体快速联检方法的建立与应用

Development and Application of A New Immunoassay for Rapid Detection of Both Mycobacterium tuberculosis and Schistosoma japonicum

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【作者】 顾炜峰王有芳王越罗学辉汤益干小仙

【Author】 GU Wei-feng, WANG You-fang, WANG Yue, LUO Xue-hui, TANG Yi, GAN Xiao-xian (Yuyao Hygiene School of Zhejiang Province, Yuyao 315400, China)

【机构】 浙江省余姚市卫生进修学校浙江省医学科学院寄生虫研究所浙江省余姚市疾病预防控制中心

【摘要】 目的创建一种新的结核、血吸虫抗体快速联合检测方法,并评价其在疾病预防中的潜在应用价值。方法以血吸虫虫卵可溶性抗原(SEA)和结核分枝杆菌脂阿拉伯甘露糖(LAM)作为检测用抗原,分别点样在硝酸纤维素膜上,以胶体金-蛋白A结合物为检测标记物,采用自行设计的垂直流检测装置为抗原抗体反应体系,建立结核抗体、血吸虫抗体快速联合检测的金标免疫渗滤法(TB/Sj-DIGFA);用TB/Sj-DIGFA检测肺结核病人、血吸虫病人、健康人及流动人口血清样本,以敏感性、特异性等指标考核其诊断价值及应用价值。结果TB/Sj-DIGFA检测36份肺结核病人血清和40份血吸虫病人血清的敏感性分别为94.4%(34/36)和100%(40/40);检测50人份健康人血清,对结核病和血吸虫病的特异性分别为80.0%(40/50)和100%(50/50)。应用新创建的TB/Sj-DIGFA方法调查3120流动人口,查出结核抗体阳性675份,阳性率为21.6%(675/3120),血吸虫抗体阳性38人份,阳性率为1.22%(38/3120)。结论TB/Sj-DIGFA检测结核和血吸虫抗体具有较高的敏感性和特异性。该方法一次操作可同时完成结核和血吸虫2项特异性抗体测定,检测效率高,可节省大量人力和物力。

【Abstract】 Objective To develop a new immunoassay for rapid detection of both M. tuberculosis and S. japonicum, and to evaluate its potential for control and prevention of tuberculosis and schistosomiasis. Method Lipoarabinomannan (LAM) of M. tuberculosis and soluble egg antigens of S. japonicum (SEA) were immobilized onto the nitrocellulose (NC) membrane. Colloidal gold-conjugated Protein-A was used as the detection marker. A specially designed home made vertical flow device (TB / Sj-DIGFA) was used in this study. Sera from infected and healthy migrant workers in Yuyao town and other provinces were then tested for the sensitivity and specificity of this system. Result Sensitivity of this new immunoassay was 94.4% (34 / 36) and 100% (40 / 40) for tuberculosis and schistosomiasis, respectively. The specificity was 80.0% (40 / 50) and 100% (50 / 50) for tuberculosis and schistosomiasis, respectively. 21.6% (675 / 3120) of the migrant workers had anti-TB antibody and 1.22% (38 / 3120) had anti-Schistosome antibody. Conclusion TB/Sj-DIGFA is a useful system with high sensitivity and high specificity. It can simultaneously detect antibodies to M. tuberculosis and S. japonicum. The operation is less time consuming and the operation is cost effective.

【基金】 浙江省钱江人才计划资助项目(No.2006R10015)
  • 【文献出处】 热带医学杂志 ,Journal of Tropical Medicine , 编辑部邮箱 ,2009年06期
  • 【分类号】R446.62
  • 【下载频次】33
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