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Archives of Virology

, Volume 164, Issue 3, pp 691–697 | Cite as

Development of a sensitive and reliable reverse transcription droplet digital PCR assay for the detection of citrus yellow vein clearing virus

  • Yingjie Liu
  • Yingli Wang
  • Qin Wang
  • Yanhui Zhang
  • Wanxia Shen
  • Ruhui Li
  • Mengji Cao
  • Lei Chen
  • Xue Li
  • Changyong ZhouEmail author
  • Yan ZhouEmail author
Original Article
  • 229 Downloads

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.

Notes

Funding

This work was supported in part by the Intergovernmental International Science, Technology and Innovation (STI) Collaboration Key Project of China’s National Key R&D Programme (NKP), The People’s Republic of China ministry of science and technology(CN) (2017YFE0110900), Overseas Expertise Introduction Project for Discipline Innovation (111 Center), Ministry of Education of the People’s Republic of China(CN) (B18044), Chongqing Research Program of Basic Research and Frontier Technology (cstc2015jcyjBX0043, cstc2017jcyjAX0150) and Fundamental Research Funds for the Central Universities, Ministry of Education of the People’s Republic of China (CN) (XDJK2018AA002).

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

705_2018_4123_MOESM1_ESM.docx (210 kb)
Supplementary material 1 (DOCX 210 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Yingjie Liu
    • 1
    • 2
  • Yingli Wang
    • 1
    • 2
  • Qin Wang
    • 1
    • 2
  • Yanhui Zhang
    • 1
    • 2
  • Wanxia Shen
    • 1
  • Ruhui Li
    • 3
  • Mengji Cao
    • 1
    • 2
  • Lei Chen
    • 4
  • Xue Li
    • 4
  • Changyong Zhou
    • 1
    • 2
    Email author
  • Yan Zhou
    • 1
    • 2
    Email author
  1. 1.National Citrus Engineering Research Center, Citrus Research InstituteSouthwest UniversityChongqingChina
  2. 2.Academy of Agricultural Sciences, Southwest UniversityChongqingChina
  3. 3.USDA-ARS, National Germplasm Resources LaboratoryBeltsvilleUSA
  4. 4.Xinping Yi-Dai Autonomous County Agricultural BureauYuxiChina

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