CAJ | 학술논문

目的：建立WHO推荐的检测人血清抗肺炎链球菌（ Streptococcus pneumoniae）荚膜多糖抗体IgG定量ELISA（Pn PS ELISA）。方法依照WHO推荐的Pn PS ELISA标准操作要求，优化荚膜多糖的包被浓度，筛选合格的ELISA板和二抗；在检测WHO参考实验室提供的质量控制血清验证实验条件的基础上，分别在WHO参考实验室和兰州生物制品研究所有限责任公司（ LIBP）实验室检测16份由LIBP制备的质控血清中抗13种肺炎链球菌血清型（1、3、4、5、6A、6B、7F、9V、14、18C、19A、19F、23F）的荚膜多糖IgG抗体浓度。同时，在LIBP 实验室建立了实验室内部质控血清907的检测范围，并评价了LIBP实验室实验间检测值的精确度。结果在WHO参考实验室和LIBP实验室分别检测的16份LIBP质控血清的抗13种肺炎链球菌血清型荚膜多糖IgG抗体浓度具有良好的相关关系（slope＝0．94，r＝0．97， P＜0．05）。 LIBP实验室的检测值有81％落在WHO参考实验检测值±40％范围内，符合评价实验室之间所用方法的75％的数据在±40％的检测值范围内的标准。建立了LIBP实验室内部质控血清907的抗13种肺炎链球菌血清型荚膜多糖抗体IgG的检测值范围。 LIBP实验室质控血清的各血清型IgG抗体浓度的实验间检测值的变异系数（ CV）均＜30％。结论 LIBP实验室成功建立了WHO推荐的检测人血清中抗肺炎链球菌荚膜多糖抗体IgG定量ELISA方法，并确定了LIBP 实验室日常用质控血清907的检测值范围。
목적：건립WHO추천적검측인혈청항폐염련구균（ Streptococcus pneumoniae）협막다당항체IgG정량ELISA（Pn PS ELISA）。방법의조WHO추천적Pn PS ELISA표준조작요구，우화협막다당적포피농도，사선합격적ELISA판화이항；재검측WHO삼고실험실제공적질량공제혈청험증실험조건적기출상，분별재WHO삼고실험실화란주생물제품연구소유한책임공사（ LIBP）실험실검측16빈유LIBP제비적질공혈청중항13충폐염련구균혈청형（1、3、4、5、6A、6B、7F、9V、14、18C、19A、19F、23F）적협막다당IgG항체농도。동시，재LIBP 실험실건립료실험실내부질공혈청907적검측범위，병평개료LIBP실험실실험간검측치적정학도。결과재WHO삼고실험실화LIBP실험실분별검측적16빈LIBP질공혈청적항13충폐염련구균혈청형협막다당IgG항체농도구유량호적상관관계（slope＝0．94，r＝0．97， P＜0．05）。 LIBP실험실적검측치유81％락재WHO삼고실험검측치±40％범위내，부합평개실험실지간소용방법적75％적수거재±40％적검측치범위내적표준。건립료LIBP실험실내부질공혈청907적항13충폐염련구균혈청형협막다당항체IgG적검측치범위。 LIBP실험실질공혈청적각혈청형IgG항체농도적실험간검측치적변이계수（ CV）균＜30％。결론 LIBP실험실성공건립료WHO추천적검측인혈청중항폐염련구균협막다당항체IgG정량ELISA방법，병학정료LIBP 실험실일상용질공혈청907적검측치범위。
Objective To establish a standardized quantitative enzyme-linked immunosorbent as-say ( ELISA) recommended by WHO for the detection of human IgG antibodies specific for Streptococcus pneumoniae capsular polysaccharides ( Pn PS ELISA ) .Methods According to the WHO recommended standard Pn PS ELISA protocol , capsular polysaccharide concentrations of 13 serotypes (1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 23F) of Streptococcus pneumonia for coating were optimized;the ELISA plates and AP conjugated goat anti-human IgG antibody for the detection were experimentally selected .Using the established assay parameters assured by testing quality control standards ( QCs) provided by WHO refer-ence laboratory , a panel of 16 LIBP ( Lanzhou Institute of Biological Products Co .Ltd.) QCs were measured for comparison analysis between WHO reference laboratory and LIBP laboratory .In the meantime , the range of IgG concentrations of an internal QC panel 907 for 13 serotypes of pneumococcal capsular polysaccharide were established for routine QC work in LIBP laboratory , and inter-assay precision within LIBP laboratory was assessed as well .Results The standardized Pn PS ELISA assay established in LIBP laboratory met the criteria required by WHO .There was a high correlation between the data collected by WHO reference labora -tory and LIBP laboratory (slope=0.94, coefficient of correlation r=0.97, P<0.05).Eighty one percent of IgG concentrations measured by LIBP laboratory were within the range of ±40%of those measured by WHO reference laboratory , which met the criteria of 75%data falling within the ambit of ±40%of assigned values commonly used for comparison .The ranges of IgG concentration in LIBP QC 907 for 13 serotypes had been established.The inter-assay precision in LIBP laboratory was high with coefficiency of variation ( CV) less than 30%.Conclusion LIBP laboratory has successfully established the standardized ELISA recommended by WHO for quantitative detection of human IgG antibodies against pneumococcal capsular polysaccharides (Pn PS ELISA).The range of IgG concentrations in LIBP QC 907 for 13 serotypes of pneumococcal capsular polysaccharide are also established for routine quality-control practice .