{"title":"Development of a SYBR Green Real-Time PCR method for the detection of Coffea arabica and C. canephora using chloroplast genes","authors":"Ju Hee Kim, Cheol Min Kim, Cheol Seong Jang","doi":"10.1007/s10068-024-01636-7","DOIUrl":null,"url":null,"abstract":"<p><i>Coffea arabica</i> (Arabica) and <i>C. canephora</i> (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.</p>","PeriodicalId":566,"journal":{"name":"Food Science and Biotechnology","volume":"7 1","pages":""},"PeriodicalIF":2.4000,"publicationDate":"2024-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Food Science and Biotechnology","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1007/s10068-024-01636-7","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"FOOD SCIENCE & TECHNOLOGY","Score":null,"Total":0}
引用次数: 0
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.
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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