抗钙调素自身抗体测定方法的建立及应用

Determination of anti一calmodulin autoantibody in the patients with autoimmune dlseases

自人脑提取钙调素（ＣａＭ），制备兔抗人ＣａＭ抗体，建立了检测抗ＣａＭ自身抗体的间接酶联免疫吸附试验方法，批内、批间误差变异系数均＜９．０％，抗ＣａＭ阳性血清用ＣａＭ吸收后吸光度值下降９０％。抗ＣａＭ抗体活性用Ｕ／ｍｌ表示，正常对照（ｎ＝１００）为７．５６±２．６８Ｕ／ｍｌ，以ｘ＋２ｓ为正常上限，＞１２．９２Ｕ／ｍｌ为阳性。各种自身免疫病患者抗ＣａＭ的水平与阳性率依次为系统性红斑狼疮（ｎ＝３２）：２６１５±１１．７Ｕ／ｍｌ，５１．１％；类风湿性关节炎（ｎ＝２８）：１３．６１±９．７Ｕ／ｍｌ，２１．４％；桥本甲状腺炎（ｎ＝２５）：５９．０７±９８．４Ｕ／ｍｌ，６０．０％；甲状腺机能亢进（ｎ＝３４）：３７．７６±８４．７Ｕ／ｍｌ，４７．１％；肝炎（ｎ＝４３）：２３．１７±６．７Ｕ／ｍｌ，４４．２％。均显著高于正常对照（Ｐ＜０．０１或Ｐ＜０．０５）。抗ＣａＭ抗体与抗核抗体常同时存在，但与抗甲状腺球蛋白、抗甲状腺微粒体自身抗体无显著相关关系（Ｐ＞０．０５）。

he activities of anti-calmodulin autoantibodies(anti-CaM)in the sera were detennhed by an ELISA technique in which CaM purified from human brain was used as coating antigen and rabbit anti一CaM antibody as positive control.The activities of positive sera(both rabbit and human anti一CaM antibodies)were greatly decreased by 90% or more when purified CaM(125mg/L)was ad-ded in the positive sera.The coefficients of intra一and interassay variation in determination of anti一CaM were both less than 90%,The results were expressed as arbitrary unit(U/ml)The levels of anti-CaM(x±s)in the 100 normal controls Were 7.56±2.68U /ml and the samples with values more than 12.92U / ml(x+2s)were regarded as positive The anti一CaM activities in the patients with systemic lupus erythematosus(SLE)(n= 32),rheumatoid arthritis(RA)(n=28),Hashimoto's thyroiditis(HT)(n=25),hyperthyroidism(n=34),and hepatitis(n=43) were 26.15±11.69U/ml,13.61±9.66U/ml,5907±98.45U/ml.37.76±84.66U/ml,and 23.17±6.69U/ml respeclively.Both the anti一CaM activities and anti一CaM positive rates in these patients were significantly higher than in the normal controls There was a significant correlation between the presence of anti一CaM antibody and anti一nuclear antibody No significant correlations were found between anti一CaM and anti一thyroglobulin or anti一thvromicrosome autoantibodies.