双抗体夹心ELISA检测生殖支原体抗原方法的初步建立及临床应用  

Establishment of antibody-sandwich ELISA method for detection of Mycoplasma genitalium antigen and its clinical application

作　　者：刘春霞[1] 辛颜彬[2] 杨雪芬[1] 魏云玲[2] 朱舜亚[2] 

机构地区：[1]张家口医学院,张家口075000 [2]军事医学科学院微生物流行病研究所,北京100071

出　　处：《军事医学科学院院刊》2002年第2期127-129,共3页Bulletin of the Academy of Military Medical Sciences

摘　　要：目的 :建立检测生殖支原体 (Mycoplasmagenitalium ,Mg)抗原的双抗夹心ELISA方法 ,探讨其临床应用的可行性。方法 :采用不同株Mg单抗用ELISA双抗夹心法检测Mg抗原 ,摸索该方法检测Mg抗原的最佳实验条件。对Mg标准株及 10 8份临床标本进行平行测定 ,观察双抗夹心ELISA方法的敏感性和特异性及该方法与PCR检测方法的符合率。结果 :包被单抗量为 2 0 μg ml,酶标记单抗 1∶2 0 0稀释时 ,P N值最大 ;检测Mg抗原敏感度达 2 μg ml;10 8份临床标本PCR检测阳性率 8.2 6 % (10 10 8) ;双抗夹心ELISA法检测阳性率 7.4 1% (8 10 8) ,两者符合率达 89.71%。结论 :建立的双抗夹心ELISA法检测Mg抗原具有快速、敏感、经济。Objective:To establish antibody_sandwich ELISA method for detection of Mycoplasma genitalium (Mg) antigen and evaluate its value for clinical application.Methods:Employing different Mg strains and monoclonal antibodies specific to Mg antigens,a new antibody_sandwich ELISA method for Mg antigen detection was established. Standard Mg strains and 108 clinical specimens were examined by this method and PCR technique parallelly in order to observe sensitivity and specificity of this method and coincidence rate between antibody_ELISA method and PCR detection. Results:In antibody_sandwich ELISA method, the optimal working concentration of capture McAb (1E12+4E7) was about 20*!μg/ml, while the best enzyme_conjugated McAb concentration(3B7) was 1∶200.The minimal Mg antigen which could be detected by this method was 2*!μg/ml.Of the 108 clinical samples examined, 10 were positive with PCR assay and 8 positive with the newly established antibody method,with the coincidence rate of these two methods being 89.71% .Conclusions:The new Mg antigen detection method has very good specificity and sensitivity,with the characters including fast to get result, easy to handle, no need of high technique and instrument and low expense. It is a valuable clinical antibody_sandwich ELISA.

关 键 词：生殖支原体 单克隆抗体 ELISA 双抗夹心法 聚合酶链反应 抗原 

分 类 号：R446.6[医药卫生—诊断学]