Food Engineering Dept, Faculty of Engineering, Erciyes Univ, 38039 Kayseri, Turkey.
J Food Sci. 2012 Feb;77(2):C167-73. doi: 10.1111/j.1750-3841.2011.02536.x. Epub 2012 Feb 6.
In this study, TaqMan-based real-time Polymerase Chain Reaction (PCR) techniques were developed for the detection of chicken and turkey meat in raw and heat-treated meat mixtures. Primers and TaqMan probe sets were designed to amplify 86 bp and 136 bp fragments for the chicken and turkey species, respectively, on the mitochondrial NADH dehydrogenase subunit 2 gene. In the results, it was possible to detect each species at the level of 0.1 pg template DNA with the TaqMan probe technique without any cross-reactivity with nontarget species (bovine, ovine, donkey, pork, and horse) while the detection level was 1 pg template DNA using conventional PCR. The TaqMan probe assays used in this study allowed the detection of as little as 0.001% level of both species in the experimental meat mixtures, prepared by mixing chicken and turkey meat with beef at different levels (0.001% to 10%). In conclusion, TaqMan probe assays developed in this research are promising tools in the specific identification and sensitive quantification of meat species even in the case of heat-treated meat products, and suitable for a rapid, automated, and routine analysis.
在这项研究中，开发了基于 TaqMan 的实时聚合酶链反应 (PCR) 技术，用于检测生肉和热处理肉混合物中的鸡肉和火鸡肉。针对线粒体 NADH 脱氢酶亚单位 2 基因，设计了用于扩增鸡和火鸡种的 86 bp 和 136 bp 片段的引物和 TaqMan 探针组。结果表明，使用 TaqMan 探针技术可以在 0.1 pg 模板 DNA 水平上检测到每种物种，而与非目标物种（牛、羊、驴、猪和马）没有交叉反应，而使用常规 PCR 的检测水平为 1 pg 模板 DNA。本研究中使用的 TaqMan 探针检测方法可以在实验性肉混合物中检测到低至 0.001%的两种物种水平，这些混合物是通过将鸡肉和火鸡肉与牛肉以不同水平（0.001%至 10%）混合制备的。总之，本研究中开发的 TaqMan 探针检测方法有望成为特定识别和敏感定量检测肉物种的工具，即使是在热处理的肉类产品中，也适用于快速、自动化和常规分析。
