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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » National Germplasm Resources Laboratory » Research » Publications at this Location » Publication #359790

Research Project: Characterizing and Detecting Pathogens to Ensure Safe Exchange of Plant Germplasm

Location: National Germplasm Resources Laboratory

Title: Development of a sensitive and reliable reverse transcription-droplet digital PCR assay for the detection of Citrus yellow vein clearing virus

Author
item Liu, Yingjie - Southwest University
item Wang, Yingli - Southwest University
item Wang, Qinshen - Southwest University
item Zhang, Yanhui - Southwest University
item Shen, Wanxia - Southwest University
item Chen, Lei - Southwest University
item Li, Xue - Southwest University
item Li, Ruhui
item Zhou, Changyong - Southwest University
item Zhou, Yan - Southwest University

Submitted to: Archives of Virology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/15/2018
Publication Date: 12/11/2018
Citation: Liu, Y., Wang, Y., Wang, Q., Zhang, Y., Shen, W., Chen, L., Li, X., Li, R., Zhou, C., Zhou, Y. 2018. Development of a sensitive and reliable reverse transcription-droplet digital PCR assay for the detection of Citrus yellow vein clearing virus. Archives of Virology. https://doi.org/10.1007/s00705-018-04123-7.
DOI: https://doi.org/10.1007/s00705-018-04123-7

Interpretive Summary: Citrus is one of the most economically important fruit crops worldwide. Citrus yellow vein disease, caused by Citrus yellow vein clearing virus (CYVCV), is one of the most serious diseases affecting lemon production in China. In this study, a new technique was developed for CYVCV detection. The assay is about 100 times more sensitive than the current detection methods. The assay was reliable when used to test 83 citrus field samples. This new assay will be useful for plant quarantine and certification programs.

Technical Abstract: Citrus yellow vein disease caused by Citrus yellow vein clearing virus (CYVCV) is one of the most serious disease affecting lemon production in China. In this study, a more sensitive and reliable reverse transcription-droplet digital PCR (RT-ddPCR) assay was developed to detect and quantify CYVCV without references. Specificity of the assay was determined by the failure of amplification of other citrus viruses. Quantitative linearity, sensitivity and accuracy of RT-ddPCR for the detection of CYVCV were compared to those of real-time RT-PCR. The results showed that both methods had a high degree of linearity (R2 = 0.9776) and quantitative correlation, but RT-ddPCR was 100 times more sensitive than real-time RT-PCR. The results of both assays for 83 samples and individual arthropods demonstrated that the RT-ddPCR assay was more sensitive and reliable. The new assay has the potential for the quantitative detection of CYVCV with high sensitivity and accuracy.