Abstract
BackgroundCrimean-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 findingsAn 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/significanceThe 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.
Highlights
Crimean-Congo Haemorrhagic fever Virus (CCHFV) is an RNA virus classified in the Orthonairovirus genus, of the family Nairoviridae, within the order Bunyavirales (International Committee on Taxonomy of Viruses 2016)
The Crimean-Congo Haemorrhagic fever (CCHF) recombinase polymerase amplification assay is a new, rapid and portable diagnostic method. It has been developed for the detection of infection with CCHF virus (CCHFV), the cause of a deadly haemorrhagic disease
CCHF is transmitted by close contact and causes a virulent haemorrhagic fever in humans for which there is no effective prophylaxis or treatment: in consequence it is classified by the Advisory Committee on Dangerous Pathogens (ACDP) as a Hazard Group 4 pathogen mandating maximum microbiological containment
Summary
CCHFV is an RNA virus classified in the Orthonairovirus genus, of the family Nairoviridae, within the order Bunyavirales (International Committee on Taxonomy of Viruses 2016). It displays a high degree of sequence variability [1] with the S-segments of CCHF viruses being phylogenetically grouped into 7 clades (Europe 1, Europe 2, Africa 1, Africa 2, Africa 3, Asia 1 and Asia 2) [2][3][4]. 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
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