역전사 다중효소 등온 급속 증폭(RT-MIRA) 기술을 이용한 수박에서 오이녹반모자이크바이러스(CGMMV) 신속 진단
Fast Diagnosis of Cucumber Green Mosaic Virus (CGMMV) in Watermelon Using Reverse Transcription Multienzyme Isothermal Rapid Amplification (RT-MIRA) Technology
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초록
Watermelon (Citrullus lanatus) is a widely cultivated fruit crop known for its nutritional value and global popularity. In Korea, eleven viruses have been identified as infecting watermelon, among which cucumber green mottle mosaic virus (CGMMV) has caused considerable yield losses. In this study, a reverse transcription–multienzyme isothermal rapid amplification (RT-MIRA) assay was developed for the detection of CGMMV in watermelon plants. A specific primer set was selected based on conserved regions of the coat protein and movement protein genes. Using this primer set, RT-MIRA successfully amplified target products, which were visualized on a 2% agarose gel. The optimized RT-MIRA conditions enabled detection within a temperature range of 35-45℃, with clear bands observed in as little as 10 minutes. Field-collected watermelon samples were also tested, and CGMMV was successfully detected using the RT-MIRA assay. These results demonstrate that the RT-MIRA assay provides a rapid, sensitive, and practical method for early detection of CGMMV in watermelon, offering strong potential for use in field diagnostics.
Abstract
Watermelon (Citrullus lanatus) is a widely cultivated fruit crop known for its nutritional value and global popularity. In Korea, eleven viruses have been identified as infecting watermelon, among which cucumber green mottle mosaic virus (CGMMV) has caused considerable yield losses. In this study, a reverse transcription–multienzyme isothermal rapid amplification (RT-MIRA) assay was developed for the detection of CGMMV in watermelon plants. A specific primer set was selected based on conserved regions of the coat protein and movement protein genes. Using this primer set, RT-MIRA successfully amplified target products, which were visualized on a 2% agarose gel. The optimized RT-MIRA conditions enabled detection within a temperature range of 35-45℃, with clear bands observed in as little as 10 minutes. Field-collected watermelon samples were also tested, and CGMMV was successfully detected using the RT-MIRA assay. These results demonstrate that the RT-MIRA assay provides a rapid, sensitive, and practical method for early detection of CGMMV in watermelon, offering strong potential for use in field diagnostics.