摘要
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.
This paper reports the application of a modified SPA-ELISA method to detect the product of Thymosin α_1 gene (Tα_1) expression. By the help of the synthesized Thymosin α_1-BSA conjugate as an antigen, the antiserum against Thymosin α_1 with a titer of 1∶2048 was prepared in rabbits. Positive results were obtained in the expression product of the recombinant plasmid (PWR590-Tα_1) after cleavage and direct coating (1∶1000 dilution) and negative results were exhibited in PWR590 control. This test was also proved to be highly specific and stable as demonstrated by the blocking test and the high consistency in 100 samples tested. In addition, the modified SPA-ELISA method showed similar grade of sensitivity and specificity to that by the double antibody sandwich method
基金
本课题由国家自然科学基金资助
关键词
胸腺素
ELISA
SPA
基因表达
Enzyme-Linked Immunosobent Assay
Staphylococcal protein A
Thymosin
Gene expression
