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用于追踪致敏性食物的首个纳米片数字PCR方法:提高了芝麻检测的灵敏度。

First nanoplate digital PCR method to trace allergenic foods: Improved sensitivity for the detection of sesame.

作者信息

Villa Caterina, Costa Joana, Mafra Isabel

机构信息

REQUIMTE-LAQV, Faculdade de Farmácia, Universidade do Porto, Rua de Jorge Viterbo Ferreira, 228, Porto 4050-313, Portugal.

REQUIMTE-LAQV, Faculdade de Farmácia, Universidade do Porto, Rua de Jorge Viterbo Ferreira, 228, Porto 4050-313, Portugal.

出版信息

Food Chem. 2024 Jun 30;444:138650. doi: 10.1016/j.foodchem.2024.138650. Epub 2024 Feb 4.

Abstract

Sesame (Sesamum indicum L.) is an important allergenic food whose presence can be the cause of severe allergic reactions in sensitised individuals. In this work, nanoplate digital PCR (ndPCR) was used to develop two methods to detect trace amounts of sesame in processed foods and compared with previously proposed real-time PCR assays. Two independent ndPCR approaches were successfully advanced, achieving sensitivities of 5 and 0.1 mg/kg of sesame in dough/biscuits, targeting the CO6b-1 and ITS regions, respectively. The sensitivity using both targets was improved by one order of magnitude comparing with real-time PCR and was not affected by food processing. CO6b-1 system was not influenced by food matrix, exhibiting similar performance regardless the use of complex matrix extracts or serial diluted DNA. Herein, ndPCR was proposed for the first time for the detection of allergenic foods with the advantage of providing better performance than real-time PCR regarding sensitivity and robustness.

摘要

芝麻(Sesamum indicum L.)是一种重要的致敏性食物,其存在可能会导致致敏个体出现严重的过敏反应。在这项研究中,纳米板数字PCR(ndPCR)被用于开发两种检测加工食品中痕量芝麻的方法,并与先前提出的实时PCR检测方法进行比较。成功推进了两种独立的ndPCR方法,分别针对CO6b-1和ITS区域,在面团/饼干中检测芝麻的灵敏度达到了5和0.1 mg/kg。与实时PCR相比,使用这两个靶点的灵敏度提高了一个数量级,并且不受食品加工的影响。CO6b-1系统不受食品基质的影响,无论使用复杂基质提取物还是系列稀释的DNA,都表现出相似的性能。本文首次提出将ndPCR用于检测致敏性食物,其优点是在灵敏度和稳健性方面比实时PCR具有更好的性能。

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