一种新的实时荧光定量PCR方法,专门检测素食原料和素食虾中的甲壳类动物

G. Cottenet, C. Blancpain
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引用次数: 2

摘要

为了符合素食主义者的要求,必须避免用真正的虾和甲壳类动物污染素食产品和素食虾,甚至是为了防止强烈的过敏反应。因此,甲壳类动物的检测必须足够可靠和敏感,以保证真实性、可追溯性和食品安全。建立了一种针对甲壳类动物线粒体16S rRNA基因的实时PCR检测方法,并对其进行了优化,以避免与引物和探针的严重错配。通过对几种甲壳类动物和常见食品成分的测试,证明了该方法仅适用于甲壳类动物。为了符合欧洲素食联盟(European Vegetarian Union)设定的0.1% (w/w)的非素食污染限值,在纯素食原料和添加了甲壳类材料的纯素食虾上确定了专用的截止CT值。该方法灵敏度≤0.0005% (w/w),在含有相近数量甲壳类动物的对照品上得到进一步验证。
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A new real-time PCR assay to specifically detect crustaceans in vegan raw materials and vegan shrimps
Abstract Contamination of vegan products and vegan shrimps with real shrimps and crustaceans must be avoided to comply with vegan claims, even more for preventing strong allergenic reactions. Therefore, the detection of crustaceans must be reliable and sensitive enough for authenticity, traceability, and food safety purposes. A new real-time PCR assay was developed targeting the mitochondrial 16S rRNA gene of crustaceans and was optimised to avoid critical mismatches with primers and probe. By testing several crustacean species and common food ingredients, the method was demonstrated to be specific to crustaceans only. To comply with the limit of non-vegan contamination established at 0.1% (w/w) by the European Vegetarian Union, dedicated cut-off CT values were determined on vegan raw materials and on vegan shrimps spiked with crustacean materials. The method reached a sensitivity ≤ 0.0005% (w/w), which was further confirmed on reference materials containing a similar amount of crustacean.
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