用于检测食品中羽扇豆痕量的商业实时聚合酶链反应方法的性能评估

Performance Evaluation of a Commercial Real-Time PCR Method for the Detection of Lupin Traces in Food.

作者信息

Tramuta Clara, Decastelli Lucia, Ingravalle Francesco, Barcucci Elisa, Fragassi Sandra, Bianchi Daniela Manila

机构信息

Experimental Zooprophylactic Institute of Piedmont, Liguria and Valle d'Aosta, Italian National Reference Center for the Detection of Food Allergens and Substances Causing Food Intolerance (CReNaRiA), 10154 Turin, Italy.

Experimental Zooprophylactic Institute of Piedmont, Liguria and Valle d'Aosta, Department Biostatistics, Epidemiology and Risk Analysis (BEAR), 10154 Turin, Italy.

出版信息

Foods. 2024 Feb 17;13(4):609. doi: 10.3390/foods13040609.

DOI:10.3390/foods13040609

PMID:38397586

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10888417/

Abstract

In accordance with U.S. FDA Foods Program Regulatory Science Steering Committee guidelines, with this study, we optimized and validated a commercial real-time PCR method for the detection of low amounts of lupin in four classes of food matrices: chocolate cookies, ragù, Olivier salad, and barley and rice flour. DNA extracted from blank (true negative) samples artificially contaminated with lupin () flour at 1000 ppm underwent dilutions with the DNA extracted from the true negative samples up to 0.5 ppm. The limit of detection for real-time PCR was 0.5 ppm in the complex matrices (range, Ct 26-34), making this a specific, robust, and rapid method for lupin allergen detection and labeling. Our validation data support the suitability of this commercially available real-time PCR method for this purpose.

摘要

根据美国食品药品监督管理局（FDA）食品项目监管科学指导委员会的指导方针，通过本研究，我们优化并验证了一种商业实时聚合酶链反应（PCR）方法，用于检测四类食品基质中的低含量羽扇豆：巧克力饼干、肉酱、奥利维尔沙拉以及大麦粉和米粉。从以1000 ppm羽扇豆（）面粉人工污染的空白（真阴性）样品中提取的DNA，与从真阴性样品中提取的DNA进行稀释，直至0.5 ppm。实时PCR在复杂基质中的检测限为0.5 ppm（范围，Ct 26 - 34），这使其成为一种用于羽扇豆过敏原检测和标记的特异性、稳健且快速的方法。我们的验证数据支持这种市售实时PCR方法适用于此目的。