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Archives of Virology

, Volume 164, Issue 7, pp 1843–1850 | Cite as

Development of a duplex reverse transcription recombinase-aided amplification assay for respiratory syncytial virus incorporating an internal control

  • Juju Qi
  • Xinna Li
  • Yi Zhang
  • Xinxin Shen
  • Guowei Song
  • Jing Pan
  • Tao Fan
  • Ruihuan Wang
  • Lixin LiEmail author
  • Xuejun MaEmail author
Brief Report

Abstract

Human respiratory syncytial virus (RSV) is a common viral pathogen that causes lower respiratory tract infections in infants and children globally. In this study, we developed a duplex reverse transcription recombinase-aided amplification (duplex-rtRAA) assay containing an internal control in a single closed tube for the detection of human RSV. The internal control in the amplification effectively eliminated false-negative results and ensured the accuracy of the duplex-rtRAA system. We first developed and evaluated a universal singleplex-rtRAA assay for RSV. The sensitivity of this assay for RSV was determined as 4.4 copies per reaction, and the specificity was 100%. Next, a duplex-rtRAA assay with an internal control was established. The sensitivity of the duplex-rtRAA assay approached 5.0 copies per reaction, and no cross-reaction with other common respiratory viruses was observed. The two detection methods (singleplex-rtRAA and duplex-rtRAA) developed in this study were used to test 278 clinical specimens, and the results showed absolute consistency with RSV RT-qPCR analysis, demonstrating 100% diagnostic sensitivity and specificity. These data indicate that the duplex-rtRAA has great potential for the rapid detection of RSV with a high sensitivity.

Notes

Acknowledgements

We acknowledge the Children’s Hospital of Hebei Province, China, for providing clinical specimens.

Funding

This work was supported by grants from China Mega-Project for Infectious Disease (2018ZX10711001, 2017ZX10104001 and 2017ZX10302301-004-002). A Chinese patent is pending.

Compliance with ethical standards

Conflict of interest

All the authors approved the final manuscript and they have no conflict of interest to declare.

Ethical approval

All aspects of the study were performed in accordance with national ethics regulations and approved by the Institutional Review Boards of National Institute for Viral Disease Control and Prevention, Center for Disease Control and Prevention of China.

Supplementary material

705_2019_4230_MOESM1_ESM.doc (48 kb)
Supplementary material 1 (DOC 47 kb)
705_2019_4230_MOESM2_ESM.doc (46 kb)
Supplementary material 2 (DOC 46 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  • Juju Qi
    • 1
    • 2
  • Xinna Li
    • 2
  • Yi Zhang
    • 2
  • Xinxin Shen
    • 2
  • Guowei Song
    • 3
  • Jing Pan
    • 3
  • Tao Fan
    • 1
    • 2
  • Ruihuan Wang
    • 2
  • Lixin Li
    • 1
    • 3
    Email author
  • Xuejun Ma
    • 2
    Email author
  1. 1.Hebei Medical UniversityShijiazhuangChina
  2. 2.Key Laboratory for Medical Virology, National Health and Family Planning CommissionNational Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and PreventionBeijingChina
  3. 3.Myasthenia Gravis Research InstituteThe First Hospital of ShijiazhuangShijiazhuangChina

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