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以酶联免疫吸附试验(ELISA)的间接法来定量不育患者血清中抗精子抗体,是灵敏和特异的实验室测试法。本文旨在研究此法与常规的标准精子凝集法(SAT)和精子制动试验(SIT)作一比较,并对本法中免疫球蛋白(Ig)的特异性、靶精子的固定,实验条件的参数选择等,作了阐述和讨论。实验血清来自40位健康男女,精液标本取自292例不育患者(21～48岁),其中7例(35～48岁),为输精管结扎术后6月至10年,作为非正常对照组。血清标本,收集后1250×g离心分离,-70℃贮存。部分血清,将用于SAT和SIT。靶精子的制备:3～5位供者精液,完成常规精液分析后,置于-25℃,集中后的精液用1∶10磷酸缓冲液(PBS,含EGTA)稀释,融解成悬浮液,600×g离心25分钟沉淀精子,弃上清液,精子沉淀后,用缓冲液再悬浮,37℃再离心,600×g10分
The indirect method of enzyme-linked immunosorbent assay (ELISA) to quantify anti-sperm antibodies in infertile patients is a sensitive and specific laboratory test. This article aims to study the method compared with the conventional standard sperm agglutination (SAT) and sperm brake test (SIT), and the specificity of immunoglobulin (Ig), target sperm fixation, experimental conditions Parameter selection, made elaboration and discussion. Serum from 40 healthy men and women semen samples from 292 cases of infertility patients (21 to 48 years), of which 7 cases (35 to 48 years), vasectomy for 6 months to 10 years, as a non-normal control group . Serum samples were collected after centrifugation at 1250 × g, stored at -70 ℃. Partial serum will be used for SAT and SIT. Target sperm preparation: 3 to 5 donor semen, after completion of conventional semen analysis, was placed at -25 ℃, after concentration of semen diluted 1:10 phosphate buffer (PBS, containing EGTA), melted into a suspension of 600 × g centrifuge for 25 minutes to sediment the sperm, the supernatant was discarded, sperm sedimentation, resuspended with buffer, centrifuged again at 37 ℃, 600 × g10 points