Location: National Germplasm Resources LaboratoryTitle: Development of a sensitive and reliable reverse transcription-droplet digital polymerase chain reaction (RT-ddPCR) assay for the detection of Citrus tristeza virus
|WANG, YINGLI - Southwest University|
|WANG, QIN - Southwest University|
|YANG, ZHEN - Southwest University|
|LIU, YINGJIE - Southwest University|
|LI, JIFEN - Southwest University|
|LI, ZHENGWEN - Southwest University|
|ZHOU, YAN - Southwest University|
Submitted to: European Journal of Plant Pathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/22/2019
Publication Date: 1/15/2020
Citation: Wang, Y., Wang, Q., Yang, Z., Li, R., Liu, Y., Li, J., Li, Z., Zhou, Y. 2020. Development of a sensitive and reliable reverse transcription-droplet digital polymerase chain reaction (RT-ddPCR) assay for the detection of Citrus tristeza virus. European Journal of Plant Pathology. https://doi.org/10.1007/s10658-019-01920-x.
Interpretive Summary: Citrus is one of the most economically important fruit crops worldwide. Citrus tristeza virus (CTV) is probably the most important virus that infects citrus, causing widespread production and economic losses. In this study, a new nucleic acid amplification technique was developed for CTV detection. The assay is 100 more sensitive than the current detection methods. The assay was reliable in detecting CTV in 55 infected field samples. The new assay will be useful for industry, plant quarantine and citrus certification programs that all need to prevent the spread of this destructive virus.
Technical Abstract: Citrus tristeza virus (CTV) is one of the most important viruses infecting citrus in the world. In this study, a sensitive and reliable assay based on the reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) was developed for the quantification of CTV with references. Specificity of the assay was determined by the failure of amplification of other relevant viruses. The quantitative linearity, sensitivity and accuracy of RT-ddPCR for the CTV detection were compared to those of real-time RT-PCR. Both methods showed a high degree of linearity (R2 = 0.991) and quantitative correlation, but ddPCR was 100 times more sensitivity than real-time RT-PCR. In summary, the results demonstrated that RT-ddPCR may contribute to improve CTV diagnosis by its higher sensitivity and specificity.