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

IF 2.4 3区 农林科学 Q3 FOOD SCIENCE & TECHNOLOGY Food Science and Biotechnology Pub Date : 2024-06-25 DOI:10.1007/s10068-024-01636-7
Ju Hee Kim, Cheol Min Kim, Cheol Seong Jang
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Abstract

Coffea arabica (Arabica) and C. canephora (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.

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利用叶绿体基因开发用于检测阿拉伯咖啡和印度咖啡的 SYBR Green Real-Time PCR 方法
阿拉比卡咖啡豆(Arabica)和罗布斯塔咖啡豆(C. canephora)是全球交易的珍贵农产品。在这项研究中,我们利用叶绿体基因为阿拉比卡咖啡豆和罗布斯塔咖啡豆设计了特定的引物对,以区分和量化这两种咖啡豆。我们使用所有引物对评估了 qRT-PCR 检测的特异性、灵敏度和适用性。设计的六对引物(三对用于阿拉比卡咖啡豆,三对用于罗布斯塔咖啡豆)的相关系数高于 0.99,斜率约为 - 3.21 至 - 3.52。效率在 92.09% 到 104.79% 之间。实时定量 PCR(qPCR)测定的检测限为 0.001 ng DNA,定量检测限为 0.01%(重量比)。此外,通过分析 12 种非目标植物物种和使用 10 种商品验证引物对的实用性,证实了引物对的特异性。本研究强调了基于 SYBR 的 qPCR 检测法在检测商业咖啡产品掺假方面的有效性。
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来源期刊
Food Science and Biotechnology
Food Science and Biotechnology FOOD SCIENCE & TECHNOLOGY-
CiteScore
5.40
自引率
3.40%
发文量
174
审稿时长
2.3 months
期刊介绍: The FSB journal covers food chemistry and analysis for compositional and physiological activity changes, food hygiene and toxicology, food microbiology and biotechnology, and food engineering involved in during and after food processing through physical, chemical, and biological ways. Consumer perception and sensory evaluation on processed foods are accepted only when they are relevant to the laboratory research work. As a general rule, manuscripts dealing with analysis and efficacy of extracts from natural resources prior to the processing or without any related food processing may not be considered within the scope of the journal. The FSB journal does not deal with only local interest and a lack of significant scientific merit. The main scope of our journal is seeking for human health and wellness through constructive works and new findings in food science and biotechnology field.
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