8、10A、11A及15B型肺炎链球菌荚膜多糖血清IgG抗体定量ELISA检测方法的验证  

Validation of quantitative ELISA for serum antibodies against pneumococcal capsular polysaccharide of serotypes 8, 10A, 11A and 15B

作　　者：李红[1] 李亚南[1] 石刚 陈琼[1] 王春娥[1] 唐静[1] 陈翠萍[1] 叶强[1] LI Hong LI Ya-nan SHI Gang CHEN Qiong WANG Chun-e TANG Jing CHEN Cui-ping YE Qiang(National Institutes for Food and Drug Control, Key Laboratory of the Method and Standardization for Quality Control of Biotechnical Products, Ministry of Public Health, Beijing 100050, China)

机构地区：[1]中国食品药品检定研究院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：《中国生物制品学杂志》2017年第9期958-962,共5页Chinese Journal of Biologicals

摘　　要：目的对8、10A、11A及15B型肺炎链球菌荚膜多糖血清Ig G抗体定量ELISA检测方法进行验证。方法以第二代国际参考血清007sp为标准,检测国际参考血清89SF的8、10A、11A及15B型Ig G抗体值,绘制标准曲线,确定检测范围、线性(R^2)及最低检测限;以第二代国际参考血清007sp为标准,测定肺炎国际质控血清盘(12/278)共12份血清的各型抗体水平3次,以第一代国际参考血清89SF为标准,测定第二代国际参考血清007sp的各型抗体水平10次,计算回收率;连续测定质控血清2009S、QC5、QC6及第二代国际参考血清007sp的各型Ig G抗体各10次,计算变异系数(CV);利用质控血清920进行抑制试验,验证方法的特异性。结果国际参考血清89SF的8、10A、11A及15B型Ig G抗体的最低检测限分别为0.58、0.85、0.46及0.67 ng/ml,检测范围分别为0.15~35.60、0.13~32.50、0.05~12.70及0.17~42.40 ng/ml,线性R^2均>0.99;除10A型的准确度较低外(46.15%),8、11A及15B型均较高(分别为76.92%、84.62%及84.62%);连续10次测定质控血清2009S、QC5、QC6及国际参考血清007sp各型Ig G抗体的CV值为6.44%~18.19%;多糖抗原与血清抗体的反应具有特异性。结论该方法具有良好的准确性、精密性及特异性,可用于肺炎球菌疫苗临床血清的检测。Objective To validate the quantitative ELISA method for serum Ig G against pneumococcal capsular polysaccharide of serotypes 8, 10 A, 11 A and 15 B. Methods The contents of serum Ig G against pneumococcal capsular polysaccharide of serotypes 8, 10 A, 11 A and 15 B in international reference serum 89 SF were determined by ELISA using the second generation of international reference serum 007 sp as standard, based on which a standard curve was plotted and determined for detection range, linearity（R^2） and minimum detection limit. The antibody levels of 12（12/278）serum samples of various serotypes in international quality control serum panel for pneumococcus were determined for 3times using the second generation of international reference serum 007 sp as standard. The antibody levels against various serotypes in the second generation of international reference serum 007 sp were determined for 10 times using the first generation of international reference serum 89 SF as standard, and the recovery rates were calculated. The Ig G levels of various serotypes in quality control sera 2009 S, QC5, QC6 and the second generation of international reference serum007 sp were determined for 10 times, of which the coefficients of variation（CVs） were calculated. Inhibition test was performed with quality control serum 920 to validate the specificity of the method. Results The minimum detection limits of Ig G against pneumococcal capsular polysaccharide of serotypes 8, 10 A, 11 A and 15 B in international reference serum89 SF were 0. 58, 0. 85, 0. 46 and 0. 67 ng/ml, with the detection ranges were 0. 15 ~ 35. 60, 0. 13 ~ 32. 50, 0. 05 ~12. 70 and 0. 17 ~ 42. 40 ng/ml, respectively, with R^2 values of more than 0. 99. Except for that for serotype 10A（46. 15%）, the accuracies of the method for determination of serotypes 8, 11 A and 15 B were 76. 92%, 84. 62% and84. 62% respectively. The CVs of 10 consecutive test results of quality control sera 2009 S, QC5, QC6 and 007 sp of various serotypes were 6. 44% ~ 18. 19%. Th

关 键 词：肺炎链球菌 荚膜多糖 血清 抗体 酶联免疫吸附试验 验证 

分 类 号：R378.1[医药卫生—病原生物学]