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开发环介导等温扩增(LAMP)检测方法,用于快速灵敏地鉴定鸵鸟肉。

Development of loop-mediated isothermal amplification (LAMP) assay for rapid and sensitive identification of ostrich meat.

机构信息

Institute of Food Quality and Food Safety, University of Veterinary Medicine, Hannover, Hannover, Germany.

出版信息

PLoS One. 2014 Jun 25;9(6):e100717. doi: 10.1371/journal.pone.0100717. eCollection 2014.

Abstract

Animal species identification is one of the primary duties of official food control. Since ostrich meat is difficult to be differentiated macroscopically from beef, therefore new analytical methods are needed. To enforce labeling regulations for the authentication of ostrich meat, it might be of importance to develop and evaluate a rapid and reliable assay. In the present study, a loop-mediated isothermal amplification (LAMP) assay based on the cytochrome b gene of the mitochondrial DNA of the species Struthio camelus was developed. The LAMP assay was used in combination with a real-time fluorometer. The developed system allowed the detection of 0.01% ostrich meat products. In parallel, a direct swab method without nucleic acid extraction using the HYPLEX LPTV buffer was also evaluated. This rapid processing method allowed detection of ostrich meat without major incubation steps. In summary, the LAMP assay had excellent sensitivity and specificity for detecting ostrich meat and could provide a sampling-to-result identification-time of 15 to 20 minutes.

摘要

动物物种鉴定是官方食品控制的主要职责之一。由于鸵鸟肉在宏观上很难与牛肉区分开来,因此需要新的分析方法。为了执行鸵鸟肉鉴定的标签法规,开发和评估一种快速可靠的检测方法可能很重要。在本研究中,基于物种鸸鹋线粒体 DNA 的细胞色素 b 基因,开发了一种环介导等温扩增(LAMP)检测方法。该 LAMP 检测方法与实时荧光计结合使用。所开发的系统允许检测到 0.01%的鸵鸟肉制品。同时,还评估了一种不使用核酸提取的直接拭子方法,使用 HYPLEX LPTV 缓冲液。这种快速处理方法允许在没有主要孵育步骤的情况下检测鸵鸟肉。总的来说,LAMP 检测法对检测鸵鸟肉具有极好的灵敏度和特异性,从采样到结果的鉴定时间为 15 到 20 分钟。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/75b0/4071038/f2018c3106a6/pone.0100717.g001.jpg

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