지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2024.10
- 수록면
- 259 - 271 (13page)
- DOI
- 10.7783/KJMCS.2024.32.5.259
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초록· 키워드
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Background: In this study, a duplex probe-based reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay was used to simultaneously detect Cycas necrotic stunt virus (CNSV) and Lychnis mottle virus (LycMoV) in Paeonia lactiflora (peony) collected from various locations in South Korea.
Methods and Results: CNSV and LycMoV infections were verified by using conventional RT- PCR using gene-specific primers. A high-precision quantification duplex probe-based qPCR assay was conducted on plasmid DNAs of target viruses and optimized to select the best-performing primers characterized by their ability to yield high relative fluorescence units and low cycle thresh- old values. Stable and consistent amplification plots and standard curves were achieved with primer and probe concentration of 0.25 µM and 0.2 µM, respectively, and an annealing temperature of 57℃. RT-qPCR was used with the selected primer sets and optimum conditions to detect the total RNA of peony leaves co-infected with CNSV and LycMoV. Successful detection occurred with a slightly weak sensitivity, having a detection limit of 10<sup>-3</sup> ng/㎕.
Conclusions: The duplex probe-based RT-qPCR assay demonstrated in this study can improve the screening process for CNSV and LycMoV, leading to a reduction in the spread of these two plant viruses
Methods and Results: CNSV and LycMoV infections were verified by using conventional RT- PCR using gene-specific primers. A high-precision quantification duplex probe-based qPCR assay was conducted on plasmid DNAs of target viruses and optimized to select the best-performing primers characterized by their ability to yield high relative fluorescence units and low cycle thresh- old values. Stable and consistent amplification plots and standard curves were achieved with primer and probe concentration of 0.25 µM and 0.2 µM, respectively, and an annealing temperature of 57℃. RT-qPCR was used with the selected primer sets and optimum conditions to detect the total RNA of peony leaves co-infected with CNSV and LycMoV. Successful detection occurred with a slightly weak sensitivity, having a detection limit of 10<sup>-3</sup> ng/㎕.
Conclusions: The duplex probe-based RT-qPCR assay demonstrated in this study can improve the screening process for CNSV and LycMoV, leading to a reduction in the spread of these two plant viruses
목차
ABSTRACT
INTRODUCTION
MATERIALS and METHODS
RESULTS
DISCUSSIONS
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