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Updated: Jul 29, 2026
Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
Published on: March 30, 2015
Development of a SYBR Green Real-Time PCR method for the detection of Coffea arabica and C. canephora using
Ju Hee Kim1, Cheol Min Kim2, Cheol Seong Jang1,2
1Agriculture and Life Sciences Research Institute, Kangwon National University, Chuncheon, Republic of Korea.
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.
Supplementary Information:
The online version contains supplementary material available at 10.1007/s10068-024-01636-7.
