摘要
目的 :建立快速测定抗人卵透明带抗体 (ZPAb)的新方法。方法 :采用人卵透明带蛋白hu2p3a2 2 -176和hu2 p3b177-3 4 8肽段作为抗原 ,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ZPAb。共检测 5 0 0份不孕者血清并与酶联免疫吸附试验 (ELISA)作对比研究。结果 :人卵透明带多肽抗原与患者血清中特异性ZPAb通过渗滤在硝酸纤维素膜上反映 ,6min内即可直接观察结果。在 5 0 0份血清检测中 ,阳性率为 2 1 2 % ,与ELISA的结果基本一致 (x2 =3.6 6 ,P >0 0 5 )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ZPAb ,具有推广使用的价值。
Objective To develop a new method for rapid identification of serum antizona-pellucida antibody in infertile women. Methods With purified huzp3a 22-176 and huzp3a 177-348 peptides as antigen and colloidal gold combined goat anti-human IgG as labelled antibody, a method of dot-immunogold filtration assay (DIGFA) based on immunocolloidal gold technic was developed for identifying human ZpAb. A total of 500 serum speciments from infertile women were examined with this method and the results compared with those detected with ELISA. Results Human Zp polypeptide antigen reacted with serum ZpAb on the nitro-cellulose membrane through filtraton and the result could be read with naked eye within 6 min. In a consecutive study of 500 specimens the positive rate was 21.20%, which corresponded well to that detected with ELISA (x 2=3.66, P>0.05). The specificity was 96.1% and sensitivity 95.4%. Funther test revealed almost nicross reactivity and excellent reproducibility. The reagents involved were stable under 0～4℃ for at least 6 months. Conclusion DIGFA can be used for rapid identification of serum ZpAb and is worth popularizing.
出处
《放射免疫学杂志》
CAS
2004年第5期408-409,共2页
Journal of Radioimmanology
