免疫金银染色法在蛋白质微阵列检测TORCH感染的实验研究

Experimental study on detection of TORCH infection using protein microarrays with immuno-gold-silver staining method

目的研制基于免疫金银染色法的TORCH检测蛋白质微阵列。方法将TORCH抗原点样于已经被琼脂糖膜修饰的玻片表面,与待测血清和胶体金标记的抗人IgM(μ链)孵育,用银增强微阵列信号,肉眼观察或普通的平板扫描仪记录结果。以不同浓度的纯化人IgM制备标准曲线。结果蛋白质微阵列的灰度质和IgM的浓度呈线性关系。60份待测血清与ELISA法同时检测,诊断符合率为93.3%。结论该法具有高信息通量、低成本、制备与检测快速简单的优点。

Objective To study a protein microarray based on immuno-gold-silver staining method for the detection of TORCH infection. Methods The antigens were printed on glass slide modified with agarose films, The gold nanoparticles were introduced to the microarray by specific binding of gold-canjugated antibodies and then coupled with silver enhancement to produce black image of microarray spots, which could be easily imaged with a simple flatbed scanner or even naked eyes, Results We created the human IgM dose-response curves which had a good linear correlation between the gray level and the sample concentration. A concordance of 93.3% was obtained between protein microarray and ELISAs in the detection for sera. Conclusions The technique of protein microarray is a rapid, simple, low-cost and high-throughput approach which is displaying more practical and broad applications.