A recombinase polymerase amplification assay for rapid detection of Crimean-Congo Haemorrhagic fever Virus infection

PLoS Negl Trop Dis. 2017 Oct 13;11(10):e0006013. doi: 10.1371/journal.pntd.0006013. eCollection 2017 Oct.

Abstract

Background: Crimean-Congo Haemorrhagic fever Virus (CCHFV) is a rapidly emerging vector-borne pathogen and the cause of a virulent haemorrhagic fever affecting large parts of Europe, Africa, the Middle East and Asia.

Methodology/principle findings: An isothermal recombinase polymerase amplification (RPA) assay was successfully developed for molecular detection of CCHFV. The assay showed rapid (under 10 minutes) detection of viral extracts/synthetic virus RNA of all 7 S-segment clades of CCHFV, with high target specificity. The assay was shown to tolerate the presence of inhibitors in crude preparations of mock field samples, indicating that this assay may be suitable for use in the field with minimal sample preparation. The CCHFV RPA was successfully used to screen and detect CCHFV positives from a panel of clinical samples from Tajikistan.

Conclusions/significance: The assay is a rapid, isothermal, simple-to-perform molecular diagnostic, which can be performed on a light, portable real-time detection device. It is ideally placed therefore for use as a field-diagnostic or in-low resource laboratories, for monitoring of CCHF outbreaks at the point-of-need, such as in remote rural regions in affected countries.

MeSH terms

  • Africa / epidemiology
  • Asia / epidemiology
  • Europe / epidemiology
  • Hemorrhagic Fever Virus, Crimean-Congo / enzymology
  • Hemorrhagic Fever Virus, Crimean-Congo / genetics
  • Hemorrhagic Fever Virus, Crimean-Congo / isolation & purification*
  • Hemorrhagic Fever, Crimean / diagnosis*
  • Hemorrhagic Fever, Crimean / epidemiology
  • Hemorrhagic Fever, Crimean / virology
  • Humans
  • Middle East / epidemiology
  • Molecular Diagnostic Techniques / economics
  • Molecular Diagnostic Techniques / instrumentation
  • Molecular Diagnostic Techniques / methods*
  • Nucleic Acid Amplification Techniques / economics
  • Nucleic Acid Amplification Techniques / instrumentation
  • Nucleic Acid Amplification Techniques / methods*
  • Point-of-Care Systems*
  • RNA, Viral / analysis
  • RNA, Viral / genetics
  • Recombinases / metabolism
  • Sensitivity and Specificity
  • Tajikistan / epidemiology
  • Time Factors

Substances

  • RNA, Viral
  • Recombinases

Grants and funding

This work was supported by funding from the Cabinet Office UKBEP /Dstl—Diagnostic support to Tajikistan. The views expressed in this publication are those of the authors and do not necessarily represent the views of the funders. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.