HBsAg弱阳性质控血清基质液的稳定性比较

Comparison of the stability of matrix solution of HBsAg weakly positive quality control serum

目的 选择不同基质配制乙肝表面抗原(HBsAg)室内质控血清用于ELISA试验,寻找配制HBsAg弱阳性质控血清的稳定基质液。方法 收集HBsAg、抗-HIV、抗-HCV、梅毒螺旋体抗体均为阴性的血清混合液作为阴性血清(HBVA),乙肝五项(HBsAg,HBsAb,HBeAg,HBeAb,HBcAb)、抗-HIV、抗-HCV、梅毒螺旋体抗体均为阴性的血清混合液作为抗体阴性血清(HBV-B),以及人血白蛋白(HBV-C)、胎牛血清(HBV-D)共4种基质,按1∶1 000、1∶2 000、1∶4 000、1∶8 000、1∶16 000、1∶32 000、1∶64 000倍比稀释HBsAg阳性血清,配制成不同基质的稀释系列,用ELISA方法检测HBsAg,选取临界值的2~4倍作为目标S/CO值,比较4种基质稀释系列在4℃下放置1、3、5、7、9、11 d、1个月和2个月,目标S/CO值对应浓度的稳定性。观察3种选定较稳定基质配制的稀释系列,4℃下放置5周和-20℃下放置6个月,目标S/CO值对应浓度的变化,比较3种选定基质配制的质控血清在-20℃下放置15个月的稳定性。结果 HBV-A为基质配制的HBsAg稀释系列稳定性不佳,4℃保存1周,目标S/CO值对应浓度降低10倍。HBV-B、HBV-C和HBV-D作为基质配制的稀释系列,4℃放置5周和-20℃放置6个月,目标S/CO值对应的浓度稳定。3种选定基质配制的质控血清-20℃放置15个月,稳定性差异不显著(P>0.05),稳定性良好。结论 阴性血清不适合作为配制HBsAg弱阳性质控血清基质,抗体阴性血清、人血白蛋白和胎牛血清稳定性良好,可作为基质使用。

Objective To select different matrices to prepare hepatitis B surface antigen(HBsAg) indoor quality control serum for ELISA,and to find a stable matrix solution for the preparation of HBsAg weakly positive quality control serum. Methods A serum mixture of negative HBsAg,anti-HIV,anti-HCV and anti-treponemal was collected as negative serum(HBV-A),four matrices including negative HBV serum(HBsAg,HBsAb,HBeAg,HBeAg,HBeAb,HBcAb)and negative for anti-HIV,anti-HCV,anti-treponemal were collected as antibody-negative serum(HBV-B),as well as four matrices,human albumin(HBV-C) and fetal bovine serum(HBV-D) were used. According to 1∶1 000,1∶2 000,1 ∶4 000,1 ∶8 000,1 ∶16 000,1:32 000,1:64 000 diluted HBsAg positive serum,a dilution series of different matrices were prepared to detect HBsAg by ELISA method. 2-4 times of the cut-off value was confirmed as the target S/CO value. The four matrix dilution series stored at 4°C for 1,3,5,7,9,11 d,1 month and 2 months,target S/CO value corresponds to the stability of the concentration were compared. The dilution series prepared with three selected more stable matrices were observed for 5 weeks at 4℃ and 6 months at-20℃,and the corresponding concentration of target S/CO values was compared,and the stability of the quality control serum prepared with the three selected matrices was compared for 15 months at-20℃. Results The HBsAg dilution series prepared with HBV-A matrix had poor stability,and the concentration corresponding to the target S/CO value was reduced by 10 after when stored at 4℃ for 1 week. Dilution series prepared as substrates were placed at 4℃ for 5 weeks and-20℃ for 6 months,with stable concentrations corresponding to target S/CO values. The quality control serum prepared by the three selected matrices was placed at-20℃ for 15 months,and the stability difference was not significant(P>0.05),the stability was good. Conclusion Negative serum was not suitable as matrix for the preparation of HBsAg weakly positive quality control serum,and antibody-negative serum,human albumin and fetal bovine serum had good stability and can be used as a matrix.