人血清中肺炎链球菌荚膜多糖IgG抗体定量ELISA的初步验证  

Preliminary validation of quantitative ELISA in determination of pneumococcal antibody in human serum

作　　者：李红[1] 李亚南[1] 石刚 刘文宾[1] 陈琼[1] 王春娥[1] 陈翠萍[1] 叶强[1] 

机构地区：[1]中国食品药品检定研究院呼吸道细菌疫苗室卫生部生物技术产品检定方法及其标准化重点实验室,北京100050

出　　处：《微生物学免疫学进展》2017年第3期24-30,共7页Progress In Microbiology and Immunology

基　　金：国家高技术研究发展计划(863计划)(2012AA02A402)

摘　　要：目的对肺炎链球菌荚膜多糖Ig G抗体定量ELISA,用于人血清中12F、19A、22F及33F型Ig G抗体的检测进行初步验证。方法以不同生产企业相同型别的12F、19A、22F及33F型荚膜多糖为包被抗原,用肺炎链球菌荚膜多糖Ig G抗体定量ELISA,对人血清中12F、19A、22F及33F型Ig G抗体进行定量检测,并对该方法的线性、检测限、检测范围、准确度、精密度、特异性进行初步验证。结果该方法检测13份质控血清的12F、19A、22F及33F型Ig G抗体的范围分别是0.02~4.38 ng/m L、0.14~34.68 ng/m L、0.10~25.20 ng/m L和0.12~29.78 ng/m L,r2均>0.99,最低检测限分别为0.35 ng/m L、0.37 ng/m L、0.44 ng/m L和0.88 ng/m L。准确度为71.15%;试验间CV值均<20%;特异性均>85%。结论肺炎链球菌荚膜多糖Ig G抗体定量ELISA,用于人血清中12F、19A、22F及33F型Ig G抗体的检测,需对准确度、精密度和特异性进一步验证。Objective The quantitative ELISA was performed and preliminary validated in determination human serum IgG antibodies against pneumococcal capsular polysaccharide serotypes 12F, 19A, 22F and 33F. Methods The pneumococcal capsular polysaccharides were used as coating antigen prepared from serotypes 12F, 19A, 22F and 33F from different enter- prises, and used a quantitative ELISA to measure the human serum IgG antibody concentration. In the meantime, the linear range, r2, detection limit, accuracy, precision and specificity related to the method were preliminary verified. Results The range of the assay was 0.02 - 4.38 ng/mL, 0.14 - 34.68 ng/mL, 0.10 -25.20 ng/mL and 0.12 - 29.78 ng/ mL resepectively for 12F, 19A, 22F and 33F serotype IgG antibody, the accuracy for 4 serotypes was 71.15% , the intra coefficient of variation（ CV} was less than 20% , the precision for each serotype was more than 85% , and the r2 was more than O. 99 for all 4 serotypes. Conclusion The accuracy, precision and specificity should be further validated in the performance of the quantitative ELISA in determination human serum IgG antibodies against pneumococcal capsular polysaccharide serotypes 12F, 19A, 22F and 33F.

关 键 词：肺炎链球菌 血清 抗体 酶联免疫吸附试验 验证 

分 类 号：R392-33[医药卫生—免疫学]