The use of colorimetric loop-mediated isothermal amplification assay for naked-eye detection of bean common mosaic virus

• Published in: Physiological and molecular plant pathology 2023-05, Vol.125, p.102017, Article 102017
• Main Authors: Çeli̇k, Ali, Morca, Ali Ferhan, Emi̇rali̇oğlu, Orkun, Yeken, Mehmet Zahit, Özer, Göksel, Çiftçi, Vahdettin
• Format: Article
• Language: English
• Subjects: BCMV; Colorimetric; One-step; RT-LAMP
• ISSN: 0885-5765; 1096-1178
• DOI: 10.1016/j.pmpp.2023.102017

Bean common mosaic virus (BCMV), one of the most prevalent viral diseases of common beans, has been intensively studied since its initial identification. In this paper, a one-step colorimetric reverse-transcription loop-mediated isothermal amplification (RT-LAMP) method was performed by designing oligonucleotide primers to amplify the gene encoding the coat protein of BCMV. Positive LAMP reactions were observed with the naked eye using WarmStart 2 × Master Mix from NEB, changing from pink to yellowish if the test is positive. The sensitivity of this method was similar to that of conventional PCR. The method was tested on common bean and another virus, bean common mosaic necrosis virus (BCMNV), and it was shown to specifically detect BCMV and not produce false positive from other RNAs. The RT-LAMP test was quite reliable for the diagnosis of BCMV from infected samples, and the precise, sensitive. The cost analysis showed that the development of a less expensive and efficient RT-LAMP assay for BCMV detection is an important step in expanding testing capacity and improving access to accurate testing in resource-limited settings. The test can be utilized in laboratory and in the field for the diagnosis and monitoring of BCMV, and is the first reported use of RT-LAMP for BCMV detection. •This study focuses on the colorimetric detection of BCMV, one of the most harmful viruses of common bean.•This article is the first to describe a colorimetric, one-step RT-LAMP methodology that was established for the diagnosis of BCMV.•The sensitivity of this method was similar to that of conventional RT-PCR.•In field samples, the novel RT-LAMP assay was shown to be accurate for diagnosing BCMV.