A Multiplex RT-PCR Assay for Detecting Three Pathogens Infecting Citrus Plants

YUAN Linkai; MA Chonghuan; LI Dingshan; CHEN Zhiwei; JIANG Xiaofeng; DING Xinlun; ZHANG Jie; WU Zujian

doi:10.19303/j.issn.1008-0384.2024.03.011

Volume 39 Issue 3

Mar. 2024

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Article Navigation > Fujian Journal of Agricultural Sciences > 2024 > 39(3): 339-344

YUAN L K, MA C H, LI D S, et al. A Multiplex RT-PCR Assay for Detecting Three Pathogens Infecting Citrus Plants [J]. Fujian Journal of Agricultural Sciences，2024，39（3）：339−344 doi: 10.19303/j.issn.1008-0384.2024.03.011

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YUAN L K, MA C H, LI D S, et al. A Multiplex RT-PCR Assay for Detecting Three Pathogens Infecting Citrus Plants [J]. Fujian Journal of Agricultural Sciences，2024，39（3）：339−344 doi: 10.19303/j.issn.1008-0384.2024.03.011

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A Multiplex RT-PCR Assay for Detecting Three Pathogens Infecting Citrus Plants

doi: 10.19303/j.issn.1008-0384.2024.03.011

Institute of Plant Virology, Fujian Agriculture and Forestry University/State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Fuzhou, Fujian　350002, China

Received Date: 2023-10-25
Rev Recd Date: 2024-01-05

Available Online: 2024-05-08

Publish Date: 2024-03-28

Abstract

Abstract

Objective A multiplex RT-PCR assay to simultaneously detect citrus yellow vein clearing virus (CYVCV), citrus tristeza virus (CTV), and hop stunt viroid (HSVd) was developed. Method Multiplex RT-PCR primers were designed, and their specificity was analyzed. The optimal concentration ratio, annealing temperature and sensitivity of primers were determined. Moreover, citrus samples from Fujian region were detected by the established multiplex RT-PCR. Result The optimized concentration ratio of CYVCV-F/R, CTV-F/R, and HSVd-F/R primers was 1:1:2, and the annealing temperature at 52.9 ℃ for the assay. The assay displayed a sensitivity at 10^-2 dilution and the positive detections on CYVCV of 47.1%, on CTV of 56.7%, and on HSVd of 22.9% on 157 detected samples. Conclusion A rapid, accurate, applicable multiplex RT-PCR method for CYVCV, CTV, and HSVd detections was successfully established.
- Citrus yellow vein clearing virus,
- citrus tristeza virus,
- hop stunt viroid,
- multiplex RT-PCR

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References(29)

References

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