免疫胶体金半定量检测牛初乳IgG含量方法的建立及初步应用

Establishment of immunogold technique half quantitative mensurated IgG of bovine colostrum and application

用柠檬酸三钠还原氯金酸,制备了20nm粒径的胶体金,颗粒均匀,在检测时颜色清晰,易于辨析。胶体金标记牛标准IgG的最适稳定量为22ug/mL。标记后免疫胶体金用含有1%BSA和0.02%叠氮钠的0.01mol/L的PBS作为稳定剂,可使制备的免疫胶体金在4℃条件下保存3个月以上。制作的检测膜片可以在15min内显示结果,且结果可长期保存。用本方法和琼脂免疫单扩散法对10份牛初乳样品和6份牛初乳制品进行检测,结果显示:牛初乳及初乳制品稀释倍数与检测膜片显色同免疫单扩散法测定的结果均有很好的相关性,但灵敏度比琼脂免疫单扩散法低。本法不仅操作过程更为简单快速,不需要特殊仪器和设备;而且检测试剂的制备简单,稳定性高,更适合于作现场实验,因此本方法具有广泛的应用及推广价值。

The colloidal gold ,which diameter is about 20nm,is obtained by reducing the gold chloride with sodium citrate and coupled with standard bovine IgG so that the immuno-colloidal gold was obtained. The amount of standard bovine IgG is 22ug/mL . 0.01mol/L PBS comprises 1% bovine serum albumin,0.02% sodium azide as stabilization of immuno-colloidal gold, it makes immuno-colloidal gold be in fair preservation.The results can be read clearly in 15 minutes and it can preserve for long time.Using the Immunogold half- quantitative method and Single immunediffusion method to test 10 bovine colostrums samples and 6 bovine colostrum product samples, the result showed that there was a nice pertinency between diluted multiple of sample and tinct change of nitrocellulose membrane with label antibody and results of Single immunediffusion method. It is unsuitable for accurate quantitative test. This method is simple,rapid and convenient tool for testing the content of IgG in bovine colostrums products in production. The detected result can be visually read by eye requiring no instrumentation.So this method has comprehensive application and extensive value.