双抗原夹心胶体金免疫层析法快速检测结核分枝杆菌抗体的研究

[A study of double antigen sandwich colloidal gold immunochromatography rapid detection for Mycobacterium tuberculosis antibody].

作者信息

Fan Chao-ming, Li Wei, Fan De-li, Zheng Xiao-yin, Ding Jian-zu

机构信息

Hangzhou Normal University, Hangzhou Second People's Hospital, Hangzhou 310015, China.

出版信息

Zhonghua Jie He He Hu Xi Za Zhi. 2011 May;34(5):356-8.

PMID:21729624

Abstract

OBJECTIVE

To establish a simple, fast and accurate double antigen colloidal gold immunochromatographic technique for detecting Mycobacterium tuberculosis antibody from tuberculosis patients.

METHODS

The fusion protein ESAT-6-16-38 was constructed by using gene cloning technique for the 6, 16 and 38 kDa early secreted antigenic target from Mycobacterium tuberculosis. The ESAT-6-16-38 fusion protein was marked to colloidal gold to establish the double antigen colloidal gold immunochromatographic assay. Serum samples from 163 patients with tuberculosis, including 57 sputum-positive cases, 64 sputum-negative cases, and 42 cases with extrapulmonary tuberculosis, were collected during 2007 and 2009 from the Disease Prevention and Control Center of Deqing County. In addition, 573 controls (224 healthy volunteers, 217 patients with acute pneumonia and bronchitis, 132 patients with paragonimiasis) were recruited for comparison. Mycobacterium tuberculosis specific antibodies were detected by using immunochromatographic and protein chip technique. Detection rate was compared with Chi-square test.

RESULTS

Among the 163 tuberculosis patients, the positive rates of immunochromatographic detection and protein chip were 73.0% (120/163) and 72.4% (118/163) respectively; the difference was not statistically significant (χ()2 = 0.062, P > 0.05). Among the 573 controls, the negative rates of immunochromatographic detection and protein chip were 93.9% (538/573) and 92.0% (527/573) respectively; the difference was not statistically significant (χ()2 = 0.635, P > 0.05). There was no cross reaction in the paragonimiasis patients. The positive rate of the immunochromatographic assay was as high as 87.7% (50/57) in the sputum-positive patients.

CONCLUSIONS

The double antigen immunochromatographic technique is an easy to operate, rapid, highly sensitive, specific, and reproducible method for the detection of Mycobacterium tuberculosis antibodies.

摘要

目的

建立一种简便、快速、准确的双抗原胶体金免疫层析技术，用于检测结核病患者的结核分枝杆菌抗体。

方法

采用基因克隆技术构建结核分枝杆菌6、16和38 kDa早期分泌抗原靶标的融合蛋白ESAT-6-16-38。将ESAT-6-16-38融合蛋白标记于胶体金上，建立双抗原胶体金免疫层析法。2007年至2009年期间，从德清县疾病预防控制中心收集了163例结核病患者的血清样本，其中痰涂片阳性57例，痰涂片阴性64例，肺外结核42例。此外，招募了573名对照者（224名健康志愿者、217例急性肺炎和支气管炎患者、132例肺吸虫病患者）进行比较。采用免疫层析法和蛋白质芯片技术检测结核分枝杆菌特异性抗体。采用卡方检验比较检测率。

结果

163例结核病患者中，免疫层析法检测阳性率为73.0%（120/163），蛋白质芯片法检测阳性率为72.4%（118/163）；差异无统计学意义（χ()2 = 0.062，P > 0.05）。573名对照者中，免疫层析法检测阴性率为93.9%（538/573），蛋白质芯片法检测阴性率为92.0%（527/573）；差异无统计学意义（χ()2 = 0.635，P > 0.05）。肺吸虫病患者无交叉反应。痰涂片阳性患者免疫层析法检测阳性率高达87.7%（50/57）。