过敏原特异性IgE检测芯片的制备及初步应用

Preparation and rudimentary application of serum allergen-specific IgE detection on protein chip

目的研制一种能同时对18种过敏原特异性IgE进行检测的蛋白芯片,并探讨其在过敏性疾病诊断中的应用价值。方法选择合适的固相载体,经L16(44×23)正交法对芯片制备,并对使用条件进行优化,以Scanarray4000激光共聚扫描仪成像,Quantarray软件分析荧光强度。同时用自制芯片和IVT试剂盒对138份过敏性疾病患者血清进行对比检测分析。结果本研究的最佳试验条件:以用NC膜为固相载体,血清稀释8倍,抗原抗体反应2h,Cy3亲和素浓度为1.25μg/ml。将本研究研制的过敏原特异性IgE检测芯片与IVT试剂盒比较,阳性检出符合率为60.2%,两种方法对鸡蛋、海鱼、大豆、花生、大籽蒿花粉等过敏原的检出率无差异(P>0.05)。结论本研究建立的过敏原特异性IgE检测芯片,具有血清用量少、同时检测多项指标、操作简便、价格低廉等优点,适合在临床检测工作中推广应用。

Objective To prepare a detective kit of serum allergen-specific IgE on protein chip and use it in clinical determination. Methods Choose an appropriate solid support, optimize the preparative andapplied conditions with orthogonal design L16（4^4×2^3 ）- The fluorescence intensities were measured by usingof Scanarray 4000, the data was analyzed by Quantarray software. The protein chip and IVT kit was comparedby 138 samples. Results Choose NC membrane as solid support. Serum dilution, antigen-antibody reactiontime and Cy3-streptavidin concentration are important factors for detective results. Compared with IVT kit,our allergen chip has no difference in detection ot the mass of antigens such as egg, sea water fish, soybean,peanut, Artemisia sieversiana pollen（x^2detect P 〉0. 05 ）. Conclusions Features of this protein chip arelittle serum sample needed, multiple detective objects, simple manipulation and low-cost format. Thismethod has obvious operational signification in clinical detective field that is the tendency of diagnosticreagents in the future.