蛋白芯片联合检测多种结核抗体在结核病诊断中的应用价值

Evaluation of a Protein Microarray detecting multi-antibodies in Mycobacteria tuberculosis diagnosis

目的设计并建立一种基于结核分支杆菌表面抗原表位的蛋白芯片检测方法,用来进行临床分离样本的检测,探讨其在临床诊断中的应用价值。方法利用生物信息学方法,分析结核分枝杆菌抗原Ag85A,38 kD,CFP10,MPT51,16 kD,TB10.4和ESAT-6的表位,并克隆表达这些优势肽段,用于制备多靶点的蛋白芯片;使用该芯片对300株临床分析样本进行检测,与商品化蛋白芯片结果进行比较,分析其敏感性与特异性。结果在200例临床确诊结核病人中,商品化芯片检测敏感性为35.5%。而本文建立的蛋白芯片检测敏感性为98.5%;在100例临床检测非结核病人中,商品化芯片特异性92%,而本文建立的蛋白芯片特异性97%。结论基于优势表位的蛋白芯片敏感性及特异性均显著高于商品化试剂盒,具有较高的临床诊断价值,可提高检测阳性率。

Objective To design and establish a protein microarray detecting method based on the peptide epitopes of Mycobacteria tuberculosis,and screen clinical isolates with this method in order to evaluate the application of this protein chip in actual clinical diagnosis.Methods Using bioinformatics method,the epitopes of Mycobacteria tuberculosis,Ag85A,38 kD,CFP10,MPT51,16 kD,TB10.4 and ESAT-6,were analyzed and these peptides were cloned and expressed to make the multi-target protein microarray.300 clinical isolates were detected with this chip,and the results were compared with the commercial products to analyze the sensitivity and specificity.Results In the 200 clinical positive isolates,the sensitivity of commercial chip is 35.5%,while that of the biochip in this article is 98.5%;in the 100 negative isolates,the specificity of commercial chip is 92%,as that of our chip is 97%.Conclusion The protein microarray shows much higher sensitivity and specificity than the commercial products in detection and maintains high clinical diagnosis value of increasing the positive detection rate.