ELISA检测抗荚膜组织胞浆菌抗体方法的建立  

Establishment of ELISA for detection of Histoplasma capsulatum antibody

作　　者：王璞[1] 陈保文[2] 罗永艾[1] 徐苗[2] 王国治[2] 

机构地区：[1]重庆医科大学附属第一医院肺科,重庆400016 [2]中国药品生物制品检定所菌苗室,北京100050

出　　处：《临床检验杂志》2007年第2期87-89,共3页Chinese Journal of Clinical Laboratory Science

基　　金：国家"863"项目(2004AA2Z3452)

摘　　要：目的建立检测小鼠血清中抗荚膜组织胞浆菌抗体的ELISA方法。方法制备荚膜组织胞浆菌(HC)抗原和结核分枝杆菌(MTB)抗原免疫小鼠,通过不同免疫次数获得不同效价的小鼠抗HC血清。以荚膜组织胞浆菌蛋白提取物(P-HTPM)为包被抗原,“方阵滴定法”确定包被抗原和抗血清的最适工作浓度,建立检测抗HC抗体ELISA法。结果HC抗原免疫小鼠3次获得4份低效价抗血清,平均A值为0.345,免疫4次获得8份高效价血清,平均A值为0.991。ELISA方法中包被抗原和抗血清的最佳工作浓度为1μg?孔及1∶1000。P-HTPM和TB-PPD与各自免疫血清均呈阳性反应,A值分别为0.993±0.216和1.606±0.263,但二者之间无交叉反应。结论本文建立的ELISA方法将成为荚膜组织胞浆菌病和结核病的一种鉴别诊断方法。Objective To establish an ELISA method for detection of antibody to Histoplasma capsulatum （HC） in mice serum. Methods Immunized-sera were acquired from mice which were immunized with HC and Mycobacterium tuberculosis. P-HTPM protein purified from HC was used as antigen in ELISA. P-HTPM and immunized-sera against HC with different dilutions were added to the wells of microtiter plate to find the optimum working concentration of antigen and antibody. Result Four lower titer and 8 higher titer immunized sera were obtained respectively through three and four times immunization. The optimum working concentrations of antigen and antibody for ELISA were 1μg of P-HTPM per well and dilution of 1 ： 1 000 of serum. Mean absorbance data of Histoplasma antibody was 0. 993 ± 0. 216 in ELISA,while the mean absorbance of tuberculosis antibody was 1. 606 ± 0. 263 using TB-PPD as antigen. There was not cross reaction between the two antigens. Conclusion ELISA using P-HTPM can be used as a method for differential diagnosis between Histoplasmasis and Tuberculosis.

关 键 词：荚膜组织胞浆菌 ELISA 结核分枝杆菌 

分 类 号：R446.5[医药卫生—诊断学]