Key Laboratory of Biomarker Based Rapid-detection Technology for Food Safety of Henan Province, Xuchang University, Xuchang 461000, People's Republic of China.
School of Food and Biological Engineering, Henan University of Science and Technology, Luoyang 471000, People's Republic of China.
J Food Prot. 2022 Aug 1;85(8):1203-1209. doi: 10.4315/JFP-22-015.
Ladder-shape melting temperature isothermal amplification (LMTIA) is a newly developed technology, and the objective of this study was to establish its effectiveness for detection of duck adulteration in beef. LMTIA primers were designed with the prolactin receptor gene of Anas platyrhynchos as the target. The LMTIA reaction system was optimized, and its performance was compared with that of the loop-mediated isothermal amplification (LAMP) assay in terms of specificity, sensitivity, and limit of detection (LOD). Our results showed that the LMTIA assay was able to specifically detect 10 ng of genomic DNAs (gDNAs) of A. platyrhynchos, without detecting 10 ng of gDNAs of Bos taurus, Sus scrofa, Gallus gallus, Capra hircus, Felis catus, and Canis lupus familiaris. The sensitivity of the LMTIA assay was 1 ng of gDNAs of A. platyrhynchos; it was able to detect duck adulteration in beef with a 0.1% LOD. Although the LAMP assay could not clearly distinguish A. platyrhynchos from G. gallus, it had a sensitivity of 10 ng of gDNAs of A. platyrhynchos and a LOD of 1% duck adulteration in beef. This study may help facilitate the surveillance of commercial adulteration of beef with duck meat.
梯型熔解温度等温扩增（LMTIA）是一种新开发的技术，本研究的目的是建立其检测牛肉中掺鸭的效果。设计了以鸭催乳素受体基因为靶标的 LMTIA 引物。优化了 LMTIA 反应体系，并在特异性、灵敏度和检测限（LOD）方面将其性能与环介导等温扩增（LAMP）检测进行了比较。结果表明，LMTIA 检测法能够特异性检测 10ng 的鸭基因组 DNA（gDNA），而不能检测 10ng 的牛、猪、鸡、羊、猫和犬的 gDNA。LMTIA 检测法的灵敏度为 1ng 的鸭 gDNA，能够检测到牛肉中掺有 0.1%的鸭。虽然 LAMP 检测法不能明确区分鸭和鸡，但它对鸭 gDNA 的灵敏度为 10ng，对牛肉中掺有 1%的鸭的 LOD 为 1%。本研究可能有助于促进对牛肉中掺鸭肉的商业掺假的监测。
