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开发一种用于检测叶绿体基因并使用SYBR Green实时荧光定量PCR的方法。 （你原文中“检测的and”表述有误，推测可能是“检测……和……”，这里根据推测进行了补全翻译）

Development of a SYBR Green Real-Time PCR method for the detection of and using chloroplast genes.

作者信息

Kim Ju Hee, Kim Cheol Min, Jang Cheol Seong

机构信息

Agriculture and Life Sciences Research Institute, Kangwon National University, Chuncheon, Republic of Korea.

Plant Genomics Laboratory, Interdisciplinary Program in Smart Agriculture, Kangwon National University, Chuncheon, Republic of Korea.

出版信息

Food Sci Biotechnol. 2024 Jun 25;34(1):115-124. doi: 10.1007/s10068-024-01636-7. eCollection 2025 Jan.

DOI:10.1007/s10068-024-01636-7
PMID:39758726
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11695554/
Abstract

UNLABELLED

(Arabica) and (Robusta) are valuable agricultural products traded worldwide. In this study, we designed specific primer pairs for Arabica and Robusta using chloroplast genes to distinguish and quantify the two types of coffee beans. We assessed the specificity, sensitivity, and applicability of the qRT-PCR assay using all the primer pairs. The six designed primer pairs (three for Arabica and three for Robusta) exhibited a correlation coefficient higher than 0.99 and a slope of approximately - 3.21 to - 3.52. The efficiency ranged from 92.09 to 104.79%. The Real-Time quantitative PCR (qPCR) assay had a detection limit of 0.001 ng DNA and a quantitative detection limit of 0.01% (w/w). Additionally, the specificity of the primer pairs was confirmed by analyzing 12 non-target plant species and verifying their practicality using 10 commercials. This study highlights the effectiveness of the SYBR-based qPCR assay in detecting adulteration in commercial coffee products.

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1007/s10068-024-01636-7.

摘要

未标注

阿拉比卡咖啡（Arabica）和罗布斯塔咖啡（Robusta）是全球交易的重要农产品。在本研究中，我们利用叶绿体基因设计了针对阿拉比卡咖啡和罗布斯塔咖啡的特异性引物对，以区分和定量这两种咖啡豆。我们使用所有引物对评估了qRT-PCR检测方法的特异性、灵敏度和适用性。设计的六对引物（三对用于阿拉比卡咖啡，三对用于罗布斯塔咖啡）的相关系数高于0.99，斜率约为-3.21至-3.52。效率范围为92.09%至104.79%。实时定量PCR（qPCR）检测方法的检测限为0.001 ng DNA，定量检测限为0.01%（w/w）。此外，通过分析12种非目标植物物种并使用10种商业产品验证其实用性，确认了引物对的特异性。本研究强调了基于SYBR的qPCR检测方法在检测商业咖啡产品掺假方面的有效性。

补充信息

在线版本包含可在10.1007/s10068-024-01636-7获取的补充材料。