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使用单拷贝基因靶点通过实时聚合酶链反应进行检测。

Detection of by Real-Time Polymerase Chain Reaction With a Single-Copy Gene Target.

作者信息

Marien Aline, Sedefoglu Hamza, Dubois Benjamin, Maljean Julien, Francis Frédéric, Berben Gilbert, Guillet Stéphanie, Morin Jean-François, Fumière Olivier, Debode Frédéric

机构信息

Quality and Authentication of Agricultural Products Unit, Knowledge and Valorization of Agricultural Products Department, Walloon Agricultural Research Centre, Gembloux, Belgium.

Haute Ecole Louvain-en-Hainaut, Montignies-sur-Sambre, Belgium.

出版信息

Front Vet Sci. 2022 Mar 9;9:718806. doi: 10.3389/fvets.2022.718806. eCollection 2022.

Abstract

Use of edible insects as an alternative source of proteins in food and feed is increasing. These last years, numerous companies in Europe have started producing insects for food and feed purposes. In the European Union, the use of edible insects for human consumption falls within Regulation (EU) No. 2015/2283 on novel foods. For feed, Commission Regulation (EU) 2017/893 authorizes seven insect species as processed animal proteins for aquaculture. Methods of authentication are required to check the conformity of the products. In this study, we propose a real-time polymerase chain reaction (PCR) method for the specific detection of the lesser mealworm (), one of the species included in the shortlist of authorized insects. The selected target is the cadherin gene with a single-copy (per haploid genome) illustrated by our experimental evidence. The PCR test amplified a 134-bp fragment of the cadherin gene. The qualitative method was assessed toward several performance criteria. Specificity was checked against 54 insect species next to other animal and plant species. The sensitivity, efficiency, robustness, and transferability of the PCR assay were also successfully tested. Finally, the applicability of the test was assessed on real-life processed samples (industrial meals) of . The study also showed that there seems to be a huge confusion on the correct labeling of the marketed mealworms. We did not succeed to get samples. They all appeared to belong to the taxon.

摘要

将可食用昆虫用作食品和饲料中蛋白质的替代来源的情况正在增加。在过去几年中,欧洲许多公司已开始生产用于食品和饲料目的的昆虫。在欧盟,将可食用昆虫用于人类消费属于关于新型食品的(欧盟)第2015/2283号法规的范畴。对于饲料,欧盟委员会法规(EU)2017/893授权七种昆虫物种作为水产养殖的加工动物蛋白。需要有认证方法来检查产品的合规性。在本研究中,我们提出了一种实时聚合酶链反应(PCR)方法,用于特异性检测小黄粉虫( ),它是授权昆虫候选名单中的物种之一。所选靶标是钙黏蛋白基因,我们的实验证据表明其为单拷贝(每个单倍体基因组)。PCR测试扩增出了钙黏蛋白基因的一个134碱基对的片段。针对几个性能标准对该定性方法进行了评估。除了其他动植物物种外,还针对54种昆虫物种检查了特异性。PCR检测的灵敏度、效率、稳健性和可转移性也得到了成功测试。最后,在真实的加工样品(工业饲料)上评估了该测试的适用性。该研究还表明,在市售黄粉虫的正确标签方面似乎存在很大的混淆。我们未能获得 样品。它们似乎都属于 分类单元。

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