09CS中11个血清型肺炎球菌荚膜多糖抗体IgG含量质控检测范围的建立

Development of determination range of IgG against pneumococcal capsular polysaccharide of 11 serotypes in quality control serum 09CS

目的建立09CS中11个血清型(2、8、9N、10A、11A、12F、15B、17F、20、22F、33F)肺炎球菌(pneumococcus,Pn)荚膜多糖抗体IgG含量的质控检测范围。方法将细胞壁C多糖(CPS)分别与Pn22F、Pn25两个型别多糖混合,制备CPS+Pn22F和CPS+Pn25两种样品吸收液,稀释09CS后,ELISA法检测13价肺炎链球菌结合疫苗(pneumococcal conjugate vaccine,13PCV)的13个血清型IgG抗体水平,验证两种吸收液检测结果的一致性。将09CS作为待测血清(用CPS+Pn25稀释),第二代国际肺炎参考血清007sp为标准血清(用CPS+Pn25稀释),第一代国际参考血清89SF为质控血清(用仅含CPS的吸收液稀释),采用ELISA法连续检测待测血清52次,计算11个血清型抗体浓度平均值(GMC)、标准偏差(SD)和变异系数(CV)、U检验中99%置信区间下的正态分布范围。结果 09CS经两种吸收液吸收后,13个型别的检测结果一致性较好,r2=0. 983 5,且差异无统计学意义(P> 0. 05)。11个血清型有效定值异常率均低于20%,异常率最大的型别为Pn20,异常率为17. 3%;各型的CV为7. 55%~12. 86%,均低于15%;Pn2、Pn8、Pn9N、Pn10A、Pn11A、Pn12F、Pn15B、Pn17F、Pn20、Pn22F、Pn33F血清型荚膜多糖抗体IgG含量的质控检测范围分别为18. 56~31. 45、13. 35~26. 61、11. 90~23. 63、14. 23~25. 74、5. 96~8. 84、3. 70~6. 47、23. 89~37. 50、10. 68~16. 90、15. 97~24. 31、10. 63~17. 61、24. 24~47. 55μg/mL。结论建立了09CS中11个血清型Pn荚膜多糖抗体IgG含量的质控检测范围,可应用于Pn疫苗临床血清荚膜多糖抗体IgG含量的ELISA法检测。

Objective To develop the determination range of IgG against capsular polysaccharide of pneumococcus(Pn)of 11 serotypes(2,8,9 N,10 A,11 A,12 F,15 B,17 F,20,22 F and 33 F)in quality control serum 09 CS.Methods Cell wall C polysaccharide(CPS)was mixed with polysaccharide of serotypes of Pn22 F and Pn25 to prepare sample absorbent solutions CPS+Pn22 F and CPS+Pn25 respectively,with which 09 CS serum was diluted and determined for IgG levels of 13 serotypes in 13 valent pneumococcal conjugate vaccine(13 PCV)to verify the consistency of test results with two absorbent solutions.The 09 CS was diluted with CPS+Pn25 and determined for 52 times by ELISA serving the second generation of international reference serum 007 sp as standard serum(diluted with the absorbent solution containing only CPS)and the first generation of international reference serum 89 SF as the quality control serum.The GMCs of IgG of11 serotypes as well as the SD,CV(%)and the normal distribution range in 99%confidence interval of U test were calculated.Results The determination results of 13 serotypes of IgG in 09 CS serum were consistent(r2=0.983 5),which showed no significant difference(P>0.05).All the abnormal rates of effective calibration numbers of the 11 serotypes were less than 20%.The abnormal rate of Pn20 was the highest,which was 17.3%.The CVs of determination results of various serotypes were 7.55%~12.86%,all of which were less than 15%.The determination ranges of IgG levels for quality control of serotypes Pn2,Pn8,Pn9 N,Pn10 A,Pn11 A,Pn12 F,Pn15 B,Pn17 F,Pn20,Pn22 F and Pn33 F were 18.56~31.45,13.35~26.61,11.90~23.63,14.23~25.74,5.96~8.84,3.70~6.47,23.89~37.50,10.68~16.90,15.97~24.31,10.63~17.61 and 24.24~47.55μg/mL respectively.Conclusion The determination range of IgG against capsular polysaccharide of Pn of 11 serotypes in quality control serum 09 CS was developed,which might be used for clinical serum assay of IgG level against capsular polysaccharide in Pn vaccine.