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目的建立一种简便快速的自身免疫抗体复合斑点胶体金法。方法将7种人细胞核抗原Sm D1、Sm B′、SSA52、SSA60、SSB、U1RNP 70K、Jo-1用毕赤酵母表达纯化后,点样于硝酸纤维素膜(NC)上,以此检测血清中相应的多种自身抗体,用SPSS 11.0统计软件分析,总符合率及7个组间符合率的比较用χ~2检验。结果与欧蒙自身抗体酶联免疫吸附法(ELISA)检测为不同自身免疫性疾病阳性血清291份和正常人血清60份比对,复合斑点胶体金法阳性符合率分别为:抗Sm D1 93.7%(59/63),抗Sm B′91.7%(55/60),抗SSA52 91.0%(81/89),抗SSA60 90.5%(76/84),抗SSB 98.6%(70/71),抗U1RNP 70K 93.6%(73/78),抗Jo-1 91.1%(51/56);阴性符合率为100%(60/60);总符合率为92.8%(465/501),统计学检验均无显著性差异(P<0.05)。结论复合斑点胶体金法简便、快速、准确,可满足临床对自身免疫性疾病联合诊断的需求。
Objective To establish a simple and rapid autoimmune antibody complex spot gold method. Methods Seven kinds of human nuclear antigens Sm D1, Sm B ’, SSA52, SSA60, SSB, U1RNP 70K and Jo-1 were expressed and purified by Pichia pastoris and then spotted on nitrocellulose membrane (NC) In the corresponding number of autoantibodies, with SPSS 11.0 statistical software analysis, the total coincidence rate and the coincidence rate between the 7 groups were compared with χ ~ 2 test. Results The results of ELISA and ELISA showed that 291 serum samples were positive for different autoimmune diseases and 60 samples were normal serum samples. The coincidence rates of anti-Sm D1 93.7% (59/63), anti-Sm B’91.7% (55/60), anti-SSA52 91.0% (81/89), anti-SSA60 90.5% (76/84), anti-SSB 98.6% The positive coincidence rate was 100% (60/60) in 70K 93.6% (73/78) and 91.1% (51/56) in anti-Jo-1. The total coincidence rate was 92.8% (465/501) Significant difference (P <0.05). Conclusions The gold chelation method of complex spot colloids is simple, rapid and accurate and can meet the need of clinical diagnosis of autoimmune diseases.