可追溯性
多重聚合酶链反应
食品科学
生肉
聚合酶链反应
生物
多路复用
物种鉴定
原材料
食品安全
生物技术
计算机科学
基因
遗传学
生态学
软件工程
作者
Song Zhou,Guowei Zhong,Hanxiao Zhou,Xiaoxia Zhang,Xiaoqun Zeng,Zhen Wu,Daodong Pan,Jun He,Zhendong Cai,Qianqian Liu
标识
DOI:10.3389/fnut.2022.890537
摘要
Frequent meat frauds have become a global issue because adulteration risks the food safety, breaches market rules, and even threatens public health. Multiplex PCR is considered to be a simple, fast, and inexpensive technique that can be applied for the identification of meat products in food industries. However, relatively less is known about a multiplex PCR method authenticating seven animal species simultaneously in one reaction due to technological challenge. Through screening new species-specific primers and optimizing PCR system, a heptaplex PCR method was established, which could simultaneously detect seven meat ingredients of camel (128 bp), pigeon (157 bp), chicken (220 bp), duck (272 bp), horse (314 bp), beef (434 bp), and pork (502 bp) in a single-tube reaction. DNA sequencing solidly validated that each set of primers specifically amplified target species from total DNA mixtures of seven meat species. The developed multiplex assay was stable and sensitive enough to detect 0.01–0.025 ng DNA from various meat treatments including raw, boiled, and autoclaved meat samples or target meat content of 0.1% total meat weight, suggesting the suitability of the heptaplex PCR technique for tracing target meats with high accuracy and precision. Most importantly, a market survey validated the availability of this multiplex PCR technique in real-world meat products with a good application foreground.
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