A novel quantitative real-time PCR method for the detection of mammalian and poultry species based on a shared single-copy nuclear DNA sequence. (30th March 2021)

• Record Type: Journal Article
• Authors: Wang, Wenjun; Fu, Ming; Zhang, Qingde; Zhen, Yueran; Liu, Jianjian; Xiang, Shengnan; Michal, Jennifer J.; Jiang, Zhihua; Zhou, Xiang; Liu, Bang
• Is Part Of: Food chemistry. Volume 341(2021)Part 2
• Publication Date: 2021-03-30
• Subjects: Quantitative real-time PCR -- Multi-species universal reference -- Meat adulteration -- Meat products; Food -- Analysis -- Periodicals; Food -- Composition -- Periodicals; 664
• DOI: 10.1016/j.foodchem.2020.128170
• Languages: English
• ISSNs: 0308-8146

Abstract

Highlights: A conserved single-copy sequence derived from LcoR gene in mammals and poultry was screened. Multi-species universal primers were used for quantitative detection among different species. Introduction of modified coefficient k improved accuracy of detection results. Improved performance of detection compared with method based on MSTN gene. Abstract: Accurate quantification of species fractions is critical to determine meat adulteration. This study aimed to develop a novel quantitative real-time PCR (qRT-PCR) method for detection of mammalian and poultry DNA. A shared single-copy nuclear DNA sequence derived from the first exon of the LcoR gene was identified as a multi-species universal reference for a qRT-PCR assay. The conservation and copy number of the LcoR gene were evaluated among different species. The limit of detection was 0.01 ng DNA or 0.01% meat ingredient, and the limit of quantification was 0.01 ng DNA or 0.05% meat ingredient. Both the relative error (R.E.) and relative standard deviation (R.S.D.) were ≤ 25%. Moreover, modified coefficient k was introduced into this quantitative system to improve the accuracy and reliability of results, with maximum R.E. improved from 19.43% to 16.16%. The quantitative method would contribute to fighting against meat adulteration and maintaining a fair market.