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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

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【作者】 白玲张志刚刘永明

【Author】 Bai Ling,Zhang Zhigang,Liu Yongming.(Faculty of Biotechnology, Guilin Medical College,Guilin 541004 China)

【机构】 桂林医学院生物技术学院桂林医学院生物技术学院 桂林541004桂林541004桂林541004

【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.

【关键词】 抗精子抗体酶联免疫吸附测定合成肽
【Key words】 AsAbELISAsynthetic peptide
【基金】 广西壮族自治区卫生厅计划课题 ( 9813 )
  • 【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
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