Res. Plant Dis > Volume 27(3); 2021 > Article
Research in Plant Disease 2021;27(3):120-127.
DOI: https://doi.org/10.5423/RPD.2021.27.3.120    Published online September 30, 2021.
Reverse Transcription Droplet Digital PCR을 활용한 Tomato Spotted Wilt Virus 검출 및 정량
이효정1, 박기범2, 한연수1, 정래동1 
1전남대학교 응용생물학과
2(주)인바이러스테크
 
Application of Reverse Transcription Droplet Digital PCR for Detection and Quantification of Tomato Spotted Wilt Virus
Hyo-Jeong Lee1, Ki Beom Park2, Yeon Soo Han1, Rae-Dong Jeong1 
1Department of Applied Biology, Institute of Environmentally Friendly Agriculture, Chonnam National University, Gwangju 61185, Korea
2Invirustech, Gwangju 91186, Korea
Received: August 23, 2021   Revised: September 27, 2021   Accepted: September 29, 2021
Abstract
Plant viruses cause significant yield losses, continuously compromising crop production and thus representing a serious threat to global food security. Tomato spotted wilt virus (TSWV) is the most harmful plant virus that mainly infects horticultural crops and has a wide host range. Reverse-transcription quantitative real-time PCR (RT-qPCR) has been widely used for detecting TSWV with high sensitivity, but its application is limited owing to the lack of standardization. Therefore, in this study, a sensitive and accurate reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) method was established for TSWV detection. Additionally, we compared the sensitivities of RT-qPCR and RT-ddPCR for TSWV detection. Specificity analysis of RT-ddPCR for TSWV showed no amplification for main pepper viruses and negative control. TSWV transcripts levels measured by RT-ddPCR and RT-qPCR showed a high degree of linearity; however, the former yielded results that were at least 10-fold more sensitive and detected lower TSWV copy numbers than the latter. Collectively, our findings show that RT-ddPCR provides improved analytical sensitivity and specificity for TSWV detection, making it suitable for identifying low TSWV concentrations in field samples.
Key Words: Detection, Reverse transcription droplet digital PCR, Tomato spotted wilt virus


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