基于胶乳增强免疫比浊法的食物过敏原检测试剂盒方法学的建立及临床应用评价

摘要

Objective:To establish a method to detect food allergen based on Latex-enhanced immune turbidimetry ( PETIA) and apply to clinic.Methods: The PETIA method was used for evaluating the detection system including standard curve , detection-limit,stability, intra-assay and inter-assay precision and cross-reactivity,clinical normal serum specimens and clinical allergic diseases serum specimens were measured to evaluated the perspective of the clinical application .Results: The standard curve :Y=837.1 x2-125.2x+10.036, linear range: 0.0-400 U/ml, the correlation coefficient of the standard curve was 0.996, the intrabatch and interbatch precision was<10%.The normal reference range was≤33.5 U/ml,the AUC of ROC curve was 0.957,the sensitivity was 89.61%,The specificity was 65.22%, the accuracy was 84.00%, and the positive predicted value was 89.61%, the negative predicted value was 65.22%.the test results have strong correlation with ELISA ( r=0.890 2 ) .Conclusion: The PETIA method for detecting food allergen achieved corresponding clinical application standards and may be used for the diagnosis of food allergies .%目的：建立基于胶乳增强免疫比浊法的食物过敏原检测试剂的方法并应用于临床。方法：采用胶乳增强免疫比浊法，对检测体系的标准曲线、最低检测量、稳定性、精密度、干扰实验等相关性能指标进行评价，通过检测健康体检人员及临床过敏人群血液标本并与目前应用于临床的较成熟的ELISA方法进行比对，评价胶乳增强免疫比浊法的食物过敏原检测试剂的临床应用可行性。结果：该方法的标准曲线 Y＝837．1 x2－125．2 x＋10．036，线性范围：0～400 U／ml，相关系数为0．996，最低检测限为19．600 U／ml。批内、批间精密度均＜10％。该方法的正常参考范围为＜33．5 U／ml，敏感度89．61％，特异性65．22％，准确度84．00％，阳性预示值89．61％，阴性预示值65．22％，ROC的曲线下面积（ AUC）为0．957，该方法与目前应用较广泛的ELISA法具有很强的相关性（ r＝0．8902）。结论：胶乳增强免疫比浊法的食物过敏原检测试剂符合临床诊断的相关要求，可应用于临床食物过敏性患者的诊断及疾病监测。