蛋白质芯片技术在过敏原特异性IgE检测中的初步应用

目的研制过敏原特异性IgE检测蛋白芯片,对该芯片的检测灵敏性和特异性进行评估。方法将不同浓度的过敏原提取液按一定阵列方式点样到固相载体上,经过固定,封闭,洗涤步骤,加入待检血清,血清中的抗体与基片上的抗原反应,用生物素-亲和素系统使Cy3荧光信号扩增,最后用Scanarray4000激光共聚扫描仪成像。与IVT试剂盒(EIA法)进行了对比检测。结果芯片的灵敏性和特异性较好,血清稀释128倍仍可检测到豚草特异性IgE的存在。芯片法对食入性过敏原的检测效果好,对吸入性过敏原的检测效果比EIA法差。结论过敏原特异性IgE抗体检测芯片,具有样本血清量少,操作简便,价格低廉等优点,具有较强的临床检测使用价值,是未来诊断试剂的发展方向。

Diagnosis of Serum Allergen-Specific IgE on Protein Chip

Objective Prepare a detective kit of serum allergen-specific IgE on protein chip and estimate the sensitivity and specificity.Methods Print allergen extracts on solid support with an ordered array. After immobilization ,blocking and washing , the membrane was incubated with human serum samples and developed antibody-antigen compounds .With the Biotin-Streptavidin system,Cy3 signals were amplified .The fluorescence intensities were measured at each spot using Scanarray 4000 . Prepare the detected results with IVT kit.Results Sensitivity and specificity of protein chip were measured,even 128-fold diluted serum could detect the existence of ragweed-specific IgE.Compared with food allergies,Pollen allergies got more false-negative results.Conclusion A desirable feature in this new diagnosis test is high-throughput ,low-cost format and little serum sample needed. Protein Chip is an useful and effective method for the measurement of multiple serum allergen-specific IgE in allergen field. More and more application to other clinical fields might be possible.