液相蛋白芯片法与BAS-ELISA法检测病原抗体的方法比较

A Comparison of Liquid Protein Chip and Biotin -avidin System -ELISA for Quantification of Immunoglobulins

〔目的〕分别建立流感病毒、禽流感病毒、鼠疫耶尔森菌、SARS病毒、结核分支杆菌等发热病原抗体的液相蛋白芯片定量检测方法,比较液相蛋白芯片方法与生物素—亲和素系统(BAS-ELISA)方法的检测能力。〔方法〕用诊断抗原包被编码微球,应用间接法检测模式,建立液相蛋白芯片定量检测病原抗体的方法,用液相蛋白芯片和BAS-ELISA方法检测相当样品并进行比较分析。〔结果〕建立的液相蛋白芯片分别检测流感、禽流感、鼠疫、SARS、结核特异性抗体,最低检测限分别为5.68ng/ml、1.50ng/ml、50.30pg/ml、105.57pg/ml、14.06pg/ml。液相蛋白芯片方法和BSA-ELISA方法的检测性能,在一定浓度范围内,对于相同样品2种方法有很好的相关性。〔结论〕与BAS-ELISA方法相比,液相蛋白芯片检测灵敏度高,动态范围宽,为同时检测多种病原抗体、任意组合检测目标物提供了平台。

Objective To develop a quantification detection of specific antibody method separately to five fevery pathogen including influenza virus,Avian influenza virus,Yersina pestis,severe acute respiratory syndrome（SARS） coronavirus and Mycobacter iumtuberculosis by liquichip protein array,and to compare the testing capabilities with the biotin-avidin system enzyme-linked immunosorbent assay（BAS-ELISA）.Method The liquichip array-based on antigen conjugated microspheres and indirect ELISA was developed for exploring the feasibility of quantitative and directly detection of specific antibody, the liquichip array was compared with a BAS-ELISA method for quantifying IgG in the same sample. Result The newly developed liquichip array have high sensitivity and wide dynamic range, the limits of detection for influenza antibody was 5.68 ng/ml; for Avian influenza antibody was 1.50 ng/ml; for Y. pestis antibody was 50.30 pg/ml; for SARS-CoV antibody was 105.57 pg/ml; and for tuberculosis antibody was 14.06 pg/ml. The liquid array had a good correlation with BSA-ELISA. Conclusion Compared with BAS-ELISA method,liquid assay has higher sensitivity and wider dynamic range, and provides a platform for simultaneous detection of pathogenic antibodies.