1. Development of a sensitive and reliable reverse transcription droplet digital PCR assay for the detection of citrus yellow vein clearing virus
- Author
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Mengji Cao, Lei Chen, Yanhui Zhang, Changyong Zhou, Xue Li, Yan Zhou, Yingjie Liu, Wanxia Shen, Qin Wang, Yingli Wang, and Ruhui Li
- Subjects
China ,Citrus ,0303 health sciences ,Reverse Transcriptase Polymerase Chain Reaction ,030306 microbiology ,General Medicine ,Biology ,Sensitivity and Specificity ,Quantitative correlation ,Virology ,Reverse transcriptase ,03 medical and health sciences ,Citrus yellow vein clearing virus ,Flexiviridae ,Digital polymerase chain reaction ,Pathogen ,Plant Diseases ,030304 developmental biology - Abstract
In 2009, a new viral disease of citrus caused by citrus yellow vein clearing virus (CYVCV) was first discovered in China. CYVCV is considered to be the most serious pathogen affecting lemon production. In this study, a sensitive and reliable reverse transcription droplet digital polymerase chain reaction (RT-ddPCR) assay was developed to detect and quantify CYVCV without references. The specificity of the assay was demonstrated by its failure to amplify other relevant citrus viruses. The quantitative linearity, sensitivity and accuracy of RT-ddPCR for detecting 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. Furthermore, RT-ddPCR was found to be 100 times more sensitive than real-time RT-PCR, and it can therefore be used to detect CYVCV in individual arthropods. In summary, the results demonstrated that the RT-ddPCR assay is a promising approach for quantitative detection of CYVCV with high precision and accuracy.
- Published
- 2018
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