免疫胶体金检测不孕症患者血清EMAb

Dot-immunogold Filtration Assay for Detection of Antiendometrial Antibody in Serum of Infertile Couples

目的 建立快速测定抗子宫内膜抗体(EMAb)的新方法。方法 采用经Sepharose4B免疫吸附柱亲和层析纯化的人子宫内膜抗原(EMAg),以胶体金颗粒结合的羊抗人IgG为标记抗体,根据免疫胶体金技术原理,建立斑点金免疫渗滤法(DIGFA)检测人血清EMAb。共检测500份不孕者血清并与酶联免疫吸附试验(ELISA)作对比研究。结果 纯化的人子宫内膜抗原与患者血清中特异性EMAb通过渗滤在硝酸纤维素膜上反应,6 min内即可直接观察结果。在500份血清检测中,阳性率为41.2％,与ELISA的结果基本一致(X2=3.66,P>0.05),符合率96.1％。特异性为95.4％,敏感性为96.3％。交叉试验与重复试验结果显示DIGFA具有较好的特异性及稳定性。结论DIGFA可快速测定EMAb,具有推广使用的价值。

Objactive To study a new method to identify antiendometrial antibody quickly in serum. Methods Purifying endometrium antigen with antiserum-Sepharose4B chemical solutions,then combining it with the nitrocellulose membrane, and taking goat anti-IgG of human combined with the golden pellets of colloidal state as marked antibody to examine antiendometrial antibody via the dot-immunogold filtration assay (DIGFA). A total of 500 clinical serum samples were ex-amined by this method and the results were compared with those detected by ELISA. Results The antiendometrial anti-body and endometrium antigen reacted on the membrane by filtration and the result could be observed with naked eyes within 6 min. The examination of 500 clinical serum samples showed that the positive rate was 41. 2%. The result of this method was in accordance with that of ELISA(X2=3. 66,P>0. 05). The specificity and sensitivity was 96. 1% and 95.4%. respectively. Cross test and repeated test showed that DIGFA had better specificity and steadiness. Conclusion The dot-immunogold filtration assay can examine antiendometrial antiboby quickly and is worth spreading.