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斑点金免疫渗滤法检测血清抗精子抗体
免疫学杂志 2001年第1期第17卷 技术与方法
作者:王虹玲 邵祝洪 胡建成 章新民
单位:王虹玲(杭州医学高等专科学校病理学教研室,浙江 杭州 310012);邵祝洪 胡建成 章新民(杭州市红十字会医院中心实验室,浙江 杭州 310003)
关键词:斑点金免疫渗滤法;人精子抗原;亲和层析;抗精子抗体
摘 要:目的 建立检测血清抗精子抗体的斑点金免疫渗滤法(dot immunogold filtration assay,DIGFA)。方法 采用经SephadexG-75-抗人全血清柱亲和层析纯化的人精子抗原和胶体金标记的SPA,根据免疫渗滤原理,建立抗精子抗体(ASAb)的斑点金免疫渗滤法检测法。结果 纯化的人精子抗原具有较好的特异性;斑点金免疫渗滤法与ELISA法比较,符合率为97.7%。特异性为96.2%(TN/TN+FP×100%),敏感性为88.9%(TP/TP+FN×100%),阳性预测值84.2%(TP/TP+FP),阴性预测值97.2%(TN/TN+FN).结论 斑点金免疫渗滤法方法检测血清抗精子抗体有较好的特异性和敏感性,重复性好,试剂稳定,操作简便快速,具有临床应用价值。
分类号:R446.61 文献标识码:A
文章编号:1000-8861(2001)01-0064-02
Dot immunogold filtration assay for detection of anti-sperm antibody in serum of infertile couples
WANG Hong-ling(Department of pathology, Hangzhou Medical Training School, Hangzhou 310012, China)
SHAO Zhu-hong(Central Laboratory Hangzhou Red Cross Hospital, Hangzhou 310003, China)
HU Jian-cheng(Central Laboratory Hangzhou Red Cross Hospital, Hangzhou 310003, China)
ZHANG Xin-min(Central Laboratory Hangzhou Red Cross Hospital, Hangzhou 310003, China)
Abstract:Objective Using dot immunogold filtration assay(DIGFA) to establish a new method for detecting anti-sperm antibody in serum of infertile couples.Methods A dot immunogold filtration assay was developed by coating purified sperm membrane antigen on nitrocellulose membrane, labeling SPA with colloidal gold, the whole procedure could be finished in 2-4 min.Results The coincidence rate of the results of simultaneous determination of serum samples with ELISA and DIGFA was 97.7%.DIGFA had an overall specificity and sensitivity of 96.2%(TN/TN+FP×100%) and 88.9%(TP/TP+FN×100%),positive predictive value and negative predictive value of 84.2% and 97.2% respectively.Conclusions Using DIGFA to detect anti-sperm antibody in human serum is more simple and rapid, and is of value for clinical application.
Keywords:DIGFA; human sperm antigen; affinity chromatography; anti-human sperm IgG
作者简介:王虹玲(1958-),男,江西赣州市人,讲师,硕士,主要从事免疫、病理和分子生物学方面的研究。Tel:(0571)8917037; E-mail:hongling021@yahoo.com参考文献:
[1]Masaru O, Morio N, Yuichi K, et al. A human sperm antigen possibly involved in binding and/or fusion with Zona-free hamster eggs[J].Fertile Steril, 1988,13(2):211-219.
[2]苗登顺,程其荣,纪凤英,等.人精子膜抗原的提纯及其在抗精抗体中的应用[J].中国实验临床免疫学杂志, 1995,7(3):33-37.
[3]Spielberg F, Kabeya CM, Ryder RW, et al. Field testing and comparative evaluation of rapid visually read screening assay for antibody to human immuno-defiency virus[J]. Lancet, 1989,1(8 638):580-584.
[4]Yakirevich E, Naot Y. Characterization of a potent sperm-agglutinating monoclonal antibody and its cognate antigens[J]. Fertil Steril, 1999,71(3):502-508.
[5]Batova IN, Ivanova MD, Mollova MV, et al. Human sperm surface glycoprotein involved in sperm-zona pellucid a interaction[J]. Int J Androl, 1998,21(3):141-153.
[6]Hiort T. Do auto-antibodies to sperm reduce fecundity? A mini-review in historical perspective[J]. Am J Reprod Immunol, 1998,40(3):215-222.
收稿日期:2000年9月11日
修稿日期:2000年10月26日
出版日期:2001年1月15日