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合成肽抗原抗精子抗体ELISA试剂盒的研究
RESEARCH OF ELISA FOR DETECTION OF ANTI-SPERM ANTIBODY WITH SYNTHETIC PEPTIDE
【摘要】 目的 :建立合成多肽抗原检测抗精子抗体 (As Ab)的酶联免疫吸附测定法 (EL ISA )。方法 :固相法合成特异性精子肽 ,包板制备检测 As Ab的 EL ISA试剂盒 ,并对该试剂盒进行方法考核。结果 :抗原包被浓度选择 0 .5 mg/ L ;检测 4 39份血清标本的 As Ab表明 ,不明原因不育患者组阳性率达 2 1.5 % ,未婚男女对照组阳性率仅为 1% ,两组对比差异有统计学意义 (P <0 .0 0 5 )。与德国 Gesellschaft试剂盒比较检测 2 6份血清标本 ,显示极好的一致性 (Kappa值 0 .77>0 .75 )。重复性试验显示其精密度较好 (CV<10 % ) ;37℃放置 8d,结果不受影响。结论 :结果表明该合成肽抗原检测抗精子抗体 EL ISA试剂盒特异性强 ,重复性好、稳定性高 ,操作简便
【Abstract】 Objective:To establish an enzyme-linked immunosorbent assay(ELISA) for the detection of anti-sperm antibody with synthetic peptide.Methods:The specific sperm peptide was synthesized by a solid-phase method. An ELISA for detecting AsAb was developed by using the peptide as the coating antigen.Result:The optimal coating concentration of the antigen was determined as 0.5 mg/L .The results of testing 439 sera with the assay showed a positive rate of infertility group was 21.5%, while unmarried youth group was 1%, (P<0.005). Comparing our assay with that of Gesellschaft in detection of 26 sera showed that both assays were very unanimous (Kappa value 0.77>0.75). Repeated test showed a low variation in detection of samples with the assay (CV<10%).The reagent was stable at 37℃ for 8 days.Conclusion:The results indicated that the ELISA for the detection of AsAb using synthetic peptide as coated antigen is specific, reproducible, stable and easy.
- 【文献出处】 广西医科大学学报 ,Journal of Guangxi Medical University , 编辑部邮箱 ,2004年04期
- 【分类号】R446.6
- 【被引频次】4
- 【下载频次】118