摘要
目的建立定量检测血清中b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)总抗体的ELISA方法,并进行初步验证。方法采用棋盘滴定法确定抗原最佳包被浓度及酶标抗体工作浓度,参考血清的线性范围及适合的底物,并验证方法的特异性、准确性及精密性。结果最佳抗原包被浓度为0.4μg/ml;最佳酶标抗体稀释度为1∶6 000;参考血清的最适线性范围为5～100 ng/ml,R2=0.997 6;选择TMB作为底物。该方法特异性较好,准确性和精密性较高。结论已成功建立了定量测定血清中Hib总抗体的ELISA方法,可逐渐替代测定血清中抗Hib IgG的方法。
Objective To develop and preliminarily verify a quantitative ELISA method for total antibody of Haemophilus influenzae type b（Hib）.Methods The coating concentration of antigen,working concentration of enzyme-labeled antibody,linear range of reference serum and substrate were optimized by chessboard titration,based on which the developed method was verified for specificity,accuracy and precision.Results The optimal coating concentration of antigen was 0.4 μg / ml,and the optimal dilution of enzyme-labeled antibody was 1 ∶ 6 000.The optimal linear range of reference serum was 5 ～ 100 ng / ml（R2 = 0.997 6）.TMB was selected as substrate.The developed method showed high specificity,accuracy and precision.Conclusion A quantitative ELISA method for total antibody of Hib was successfully developed,which might substitute to the determination of anti-Hib IgG in sera.
出处
《中国生物制品学杂志》
CAS
CSCD
2011年第8期974-976,共3页
Chinese Journal of Biologicals
