检测抗白色念珠菌烯醇化酶IgG抗体免疫胶体金层析试纸的制备

Preparation of Chromatography Test Paper for Detection of Anti-candida Albicans Enolase IgG Antibody by Using Immune Gold Colloid

目的建立一种快速检测抗白色念珠菌烯醇化酶IgG抗体的免疫胶体金层析试纸法。方法以重组烯醇化酶包被于硝酸纤维素膜上,与金标垫上包被的金标小鼠抗人IgG单克隆抗体配对,优化试纸制备条件并制备试纸条,检测人血清中的抗念珠菌烯醇化酶IgG抗体。用该方法对38例确证念珠菌血流感染者和50例健康体检者血清进行检测,分析其检测效能。结果用直径30 nm金颗粒制备的烯醇化酶IgG抗体检测试纸条,最适烯醇化酶包被浓度为1. 0 mg/ml。该试纸用于检测多种念珠菌引起的血流感染中的抗烯醇化酶IgG抗体,检测灵敏度、特异性、阳性预测值和阴性预测值分别为71. 1%,96. 0%,93. 1%和83. 4%。结论用重组白色念珠菌烯醇化酶制备的免疫胶体金层析试纸条有较好的检测效能,具有准确、快速、方便的优点,可用于人血清抗念珠菌IgG抗体的检测。

Objective To establish a preparation method of chromatography test paper for rapid detection of anti-candida albicans enolase （Eno） IgG antibody by using immune gold colloid. Methods Recombinant Eno was immobilized on nitrocellulose filter, and was matched with anti human IgG monoclonal antibodies of gold marking mouse, which immobilized on gold marking pad, and then conditions of preparing test paper were optimized to prepare test strip, and human serum anti-candida Eno IgG antibodies were detected. The method was used to detect serum samples from 38 patients with candida infection and 50 healthy people so as to analyze detection efficiency of the method. Results Gold particles of 30 nm were used for preparing test strip of detecting Eno IgG antibody, and the optimal concentration of recombinant Eno for immobilization was 1.0 mg/ml. The test paper could be used to detect anti Eno IgG antibody induced by candida in infected blood samples, and sensitivity, specificity, positive predictive value （PPV） and negative predictive value （NPV） were 71.1 %, 96.0 %, 93.1 % and 81.4 % respectively. Conclusion Immune colloidal gold chromatography test paper prepared with recombinant candida albicans enolase has good detection efficiency with accurate, quick and convenient, and therefore it can be used to detect human serum anti- candida IgG antibody.