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本文报道检测人胸腺素α_1基因表达产物的改良SPA-ELISA法.人工合成的Tα_1与BsA相偶联后,经微量免疫法获得了1∶2048滴度的抗Tα_1抗血清.本试验将人Tα_1基因的克隆株PWR590表达的裂解液,以1∶1000稀释后直接包被,经抗Tα_1抗血清、HRP-SPA及邻苯二胺底物的酶标法,成功地进行了表达产物的Tα_1检测.根据无Tα_1基因的PWR590克隆株表达的裂解液的结果为阴性,而以人工合成的Tα_1包被者为阳性;阻断试验成立,说明本试验具有较好的特异性.经100份样本的检测,均获得了一致的结果,提示本法稳定.同时我们还将其与双抗体夹心法进行了比较,结果两者亦基本一致.
In this study, a modified SPA-ELISA method was developed to detect the expression of human thymosin α_1 gene.After the synthetic Tα_1 was coupled with BsA, the 1:2048 titers of anti-Tα_1 antiserum were obtained by microimmunization.In this study, human Tα_1 The lysate of the cloned strain PWR590 was directly coated at a dilution of 1: 1000. The Tα_1 of the expressed product was successfully detected by ELISA with anti-Tα 1 antiserum, HRP-SPA and o-phenylenediamine substrate .The result of the lysate expressed by the PWR590 clones without Tα_1 gene was negative and the Tα_1-coated by human was positive.The blocking test was established to show that the test had good specificity.After 100 samples Test, have obtained consistent results, suggesting that this method is stable.At the same time, we also compared with the double antibody sandwich method, the results are basically the same.