纯芝麻酱中花生致敏蛋白Ara h2、Ara h3及芝麻蛋白2S albumim的分析鉴定比较

Analysis and Comparison of Peanut Allergen Proteins Ara h2, Ara h3 and Sesame Protein 2S albumin in Pure Sesame Paste

摘要:

针对花生致敏蛋白Ara h2、Ara h3及芝麻蛋白2S albumim，使用实时荧光定量PCR法（Real-time PCR）及酶联免疫吸附法（ELISA）对纯芝麻酱中上述蛋白进行检测。两种方法的特异性、重复性方法学验证显示均良好。验证后使用实时荧光定量PCR法对纯芝麻酱DNA进行芝麻蛋白2S albumim基因、花生蛋白Ara h2基因的检测，使用酶联免疫吸附法对花生致敏蛋白Ara h3检测。实时荧光定量PCR法检测90批样品均含有芝麻成分，CT值为22.00~33.80，其中45批次还检出花生成分，CT值为25.60~38.60；酶联免疫吸附法除检出此45批次外还另检出38批次含有花生成分。通过结果比对分析及灵敏度检测研究发现，酶联免疫吸附法检出限更低。二者检测结果虽有差异，但不矛盾，均可用于纯芝麻酱中花生源检测。高含量样品检测时可选择实时荧光定量PCR法，低含量或无法预估的样品可选择酶联免疫吸附法。目前无芝麻酱中过敏原检测相关标准，该研究可作为基础研究的可靠数据并建议监管部门尽快建立相关方法，更好的规范市场行为，保障人民的饮食安全。

Abstract:

In this study, real-time PCR（RT-PCR）and enzyme-linked immunosorbent assay (ELISA) were used to detect peanut allergenic proteins Ara h2, Ara h3 and sesame protein 2S albumin in pure sesame paste. The specificity and reproducibility of the two methods were shown to be good. After verification, real-time PCR was used to detect the sesame protein 2S Albumin gene and peanut allergen Ara h2 gene, and ELISA was used to detect the peanut allergen Ara h3 protein. The 90 batches of samples detected by real-time PCR test contained sesame components (with CT value ranging from 22.00 to 33.80), and among which 45 batches of samples contained peanut components (CT value: 25.60~38.60). Besides these 45 batches s, ELISA also allowed the detection of peanut components in another 38 batches. By comparing and analyzing the results of the two methods and through sensitivity analysis research, it was found that the ELISA method had a lower detection limit. Although the results of the two methods were different, they were not contradictory, with both suitable for the detection of peanut sources in pure sesame samples. The real-time PCR method is appropriate for the samples with a high peanut content, whilst the ELISA method can be selected for samples with a low peanut content or unpredictable samples. At present, there are no relevant standards for the detection of allergens in sesame paste. This study provides reliable data for basic research, and is the results suggest that the regulatory authorities should establish relevant methods as soon as possible to supervise market behavior and ensure people's dietary safety.