白色念珠菌烯醇化酶抗原双抗体夹心法的建立及应用  

Development and application of an double-antibody sandwich ELISA method for the detection of enolase from Candida albicans

作　　者：赵会海 贺政新 李芳 高鹏 王佳钰 王芳芳 秦立霞 ZHAO Hui-hai;HE Zheng-xin;LI Fang;GAO Peng;WANG Jia-yu;WANG Fang-fang;QIN Li-xia(Department of Clinical Laboratory,the 980th Hospital of Joint Logistics Support Force of Chinese PLA,Shijiazhuang 050082,Hebei,CHINA)

机构地区：[1]联勤保障部队第980医院检验科,河北石家庄050082

出　　处：《海南医学》2023年第24期3600-3603,共4页Hainan Medical Journal

基　　金：河北省卫健委医学科学研究重点课题计划项目(编号:20180918)。

摘　　要：目的建立定量检测白色念珠菌烯醇化酶抗原的双抗体夹心法,为后续定量检测念珠菌性阴道炎患者阴道分泌物中的烯醇化酶抗原提供方法学基础。方法用方阵滴定法确定最佳包被抗体浓度和最佳酶标二抗浓度,建立检测白色念珠菌烯醇化酶抗原的ELISA法,用该法检测白色念珠菌标准菌株SC5314培养上清中烯醇化酶抗原的浓度,初步探索应用于诊断外阴阴道念珠菌病。结果最佳包被抗Eno单抗浓度为4.15μg/mL;酶标二抗HRP标记的抗烯醇化酶多抗最佳稀释倍数为1∶1000。在第12小时时白色念珠菌培养上清中的烯醇化酶抗原开始检出,随着培养时间的延长,其上清中Eno抗原的浓度也随着逐渐增高,第48小时时达到峰值后趋于稳定。阴道分泌物上清中烯醇化酶抗原浓度随着真菌培养菌落计数的增加其浓度也在增加,有很好的相关性。结论成功建立了定量检测白色念珠菌烯醇化酶抗原的双抗体夹心ELISA方法,可应用于评价念珠菌性阴道炎患者阴道分泌物中的烯醇化酶抗原的水平。Objective To establish an double-antibody sandwich ELISA method for the detection of enolase from Candida albicans and provide the basis for the subsequent quantitative detection of enolase(Eno)antigen in vaginal secretions of patients with Candida vaginitis.Methods The optimal concentrations of coating anti-body and enzyme-labeled secondary antibody were determined with the square matrix titration.An ELISA method was established to detect the enolase antigen of Candida albicans.The concentration of enolase antigen in culture supernatant of standard strain SC5314 was detected by this method,and preliminary exploration was applied to the diagnosis of Vulvovaginitis candidiasis(VVC).Results The optimal concentration of Eno was 4.15μg/mL,and the optimal dilution ratio of HRP-labeled anti-enolase polyclonal antibody was 1∶1000.The enolase antigen in the supernatant of Candida albicans culture began to be detected at the 12th hour.With the extension of culture time,Eno antigen concentration in the supernatant increased gradually,reaching a peak at the 48th hour and then stabilizing.The concentration of enolase antigen in the supernatant of vaginal secretions increased with the increase of fungal colony count,and there was a good correlation.Conclusion A double-antibody sandwich ELISA method for detecting enolase antigen of Candida albicans was established successfully,which can be used to evaluate the level of enolase antigen in vaginal secretions of patients with Candida vaginitis.

关 键 词：念珠菌性阴道炎 白色念珠菌 烯醇化酶 双抗体夹心酶联免疫吸附试验 

分 类 号：R711.31[医药卫生—妇产科学]