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作 者:金鑫 陈红霞(指导)[1] JIN Xin;CHEN Hong-Xia(School of Life Sciences,Shanghai University,Shanghai 200444,China)
出 处:《中国免疫学杂志》2020年第16期1983-1987,共5页Chinese Journal of Immunology
摘 要:目的:利用抗免疫复合物IgG多抗体在光激化学发光(Lica)平台上建立间接法检测甲状腺过氧化物酶抗体(Anti-TPO)。方法:以人体IgG对兔子进行免疫耐受,以兔红细胞和人体血清构建免疫复合物IgG并免疫兔子。用纯化的抗免疫复合物IgG多抗和甲状腺过氧化物酶组成一步间接法,检测甲状腺过氧化物酶抗体浓度,并评估其最低检测限、回收率和批内精密度,与传统两步化学发光法(贝克曼)进行比较。结果:甲状腺过氧化物酶的检测方法的最低检测限为0.09 U/ml,平均回收率为96.26%,重复性变异系数(CV)为1.58%~1.97%,与贝克曼方法的结果的相关性r=0.989。结论:成功建立了光激化学发光定量检测Anti-TPO的方法(间接法)。该方法与传统两步间接化学发光法的相关性较好,弥补了光激化学发光平台无法使用间接法进行检测的缺陷。Objective:An indirect method for the detection of thyroid peroxidase antibodies was established on the platform of light initiated chemiluminescent immunoassay(Lica)using polyclonal antibodies of immune complexes IgG.Methods:Immuned tolerance to rabbits with human IgG,and rabbits were immunized with immune complexes IgG that were constructed with rabbit red blood cells and human serum.One-step indirect method consisting of purified polyclonal antibodies of immune complexes IgG and thyroid peroxidase was used to detect the thyroid peroxidaseautoantibodies concentration,and the limit of detection,recovery rate,intra-assay precision and comparing with the Beckman Coulter were evaluated.Results:The limit of detection of the thyroid peroxidaseautoantibodies assay was 0.09 U/ml,the average recovery was 96.26%,the repeatability coefficient of variation(CV)was 1.58%-1.97%,and the correlation with the results of Beckman method was r=0.989.Conclusion:Successsfully esatablished an indirect method by employing initiated chemiluminescent immunoassay for detection of Anti-TPO.The assay This method has a good correlation with the traditional two-step Indirect Chemiluminescence method,and makes up for the defect that initiated chemiluminescent immunoassay can not be detected by indirect method.
关 键 词:光激化学发光法 间接法 抗免疫复合物IgG多抗 甲状腺过氧化物酶抗体 微粒
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