食物过敏原特异性IgE检测方法的建立及临床应用

The establishment of a new method to detect food allergen specific IgE and its clinical application for detecting allergic diseases

目的研制一种能同时检测多种食物过敏原特异性IgE检测条，并探讨其在过敏性疾病诊断中的应用价值。方法制备常见食物过敏原的提取液，以硝酸纤维素膜（NC）膜为固相载体制备过敏原检测条，并对制备条件和检测条件进行优化。用自制检测条与德国Allergy Screen过敏原检测系统对210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为，以NC膜为固相载体，点样后室温下包被2h，2％聚乙烯醇（PVA）室温封闭1h，血清稀释10倍，抗原抗体室温下反应2h，酶标的抗人IgE抗体浓度为2μg／ml。对210份过敏性疾病患者血清进行检测，Allergy Screen过敏原检测系统检出阳性标本154份，阴性标本56份，自制检测条检出阳性标本104份，阴性标本106份。2种方法的阳性结果符合率为63．6％，阴性结果符合率为94．6％。2种方法对鸡蛋白、牛奶、花生、黄豆、螃蟹、虾等大多数过敏原的检出率差异无统计学意义（x^2值分别为2．53、2．40、2．08、2．38、0．17、1．13，P均〉0．05）。自制检测条具有较好的重复性和稳定性。结论自制多种过敏原特异性IgE检测条具有血清用量少、同时检测多种过敏原、操作简便、价格低廉等优点，且具有较强的临床检测应用价值。

Objective To prepare a detective membrane strip for detection of food allergen-specific IgE in serum samples and estimate its clinical application value in allergic diseases. Methods The crude extracts of the food allergens were prepared. Nitrocellulose membrane as the solid support was selected and the coating and the detecting conditions were optimized. The membrane strips were used to detect serum samples in 210 patients with allergic diseases and the results were compared with German Allergy Screentesting system. Results The optimal experimental conditions were as follows： The NC membrane was adopted as the solid support. After being spotted, the food allergens were incubated for 2 hours at room temperature, followed by 2% PVA blocking for 1 hour. After serum samples were diluted （1： 10） and incubated for 2 hours at room temperature, the concentration of anti-human IgE was 2 μg/ml. Compared with the German Allergy Screen-testing system, their positive detectical coincidence was 53.6% , and negative detectical coincidence was 94. 5%. The two methods had no difference in detecting the majority of food allergens such as egg white, milk, peanut, soybean, crab and shrimp （X^2 2. 53, 2.40, 2. 08, 2. 38, 0. 17, 1.13, P 〉 0. 05 ）. Conclusions The advantages of our method for detecting allergic diseases are little serum needed, multiple detective allergens, simple manipulation and low cost. This method has obvious clinical application value, which should be a new detective method for the allergic diseases with broad perspectives.