Virus Detection and Monitoring of Viral Load in Crimean-Congo Hemorrhagic Fever Virus Patients

Wölfel, Roman; Pawęska, Janusz T.; Petersen, Nadine; Grobbelaar, Antoinette A.; Leman, Patricia A.; Hewson, Roger; Georges‐Courbot, Marie‐Claude; Papa, Anna; Günther, Stephan; Drosten, Christian

doi:10.3201/eid1307.070068

Cited by 116 publications

(100 citation statements)

References 12 publications

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“…The resulting high viremia levels in the blood on days 2 and 3 postinfection are comparable to severe disease in human cases (7,36). In STAT129 mice, the liver and spleen are the major sites of replication and the only sites where prominent histopathologic changes were noticed.…”

Section: Vol 84 2010 New Animal Model For Crimean-congo Hemorrhagicmentioning

confidence: 66%

“…To determine viral titers in tissues, a 3-mm by 3-mm piece of tissue was removed during necropsy and homogenized using metal beads (Tissue Lyser, Qiagen) in RLT buffer (Qiagen), and RNA was extracted using an RNeasy kit (Qiagen) according to the manufacturer's instructions. CCHFV-specific quantitative real-time reverse transcription (RT)-PCR was performed as previously described (36). Viral titers were reported as the genome equivalence (GEQ).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Pathogenesis and Immune Response of Crimean-Congo Hemorrhagic Fever Virus in a STAT-1 Knockout Mouse Model

Bente

Alimonti

Shieh

et al. 2010

J Virol

210

242

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Tick-borne Crimean-Congo hemorrhagic fever virus (CCHFV) causes a severe hemorrhagic syndrome in humans but not in its vertebrate animal hosts. The pathogenesis of the disease is largely not understood due to the lack of an animal model. Laboratory animals typically show no overt signs of disease. Here, we describe a new small-animal model to study CCHFV pathogenesis that manifests clinical disease, similar to that seen in humans, without adaptation of the virus to the host. Our studies revealed that mice deficient in the STAT-1 signaling molecule were highly susceptible to infection, succumbing within 3 to 5 days. After CCHFV challenge, mice exhibited fever, leukopenia, thrombocytopenia, and highly elevated liver enzymes. Rapid viremic dissemination and extensive replication in visceral organs, mainly in liver and spleen, were associated with prominent histopathologic changes in these organs. Dramatically elevated proinflammatory cytokine levels were detected in the blood of the animals, suggestive of a cytokine storm. Immunologic analysis revealed delayed immune cell activation and intensive lymphocyte depletion. Furthermore, this study also demonstrated that ribavirin, a suggested treatment in human cases, protects mice from lethal CCHFV challenge. In conclusion, our data demonstrate that the interferon response is crucial in controlling CCHFV replication in this model, and this is the first study that offers an in-depth in vivo analysis of CCHFV pathophysiology. This new mouse model exhibits key features of fatal human CCHF, proves useful for the testing of therapeutic strategies, and can be used to study virus attenuation.

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Section: Vol 84 2010 New Animal Model For Crimean-congo Hemorrhagicmentioning

confidence: 66%

Section: Methodsmentioning

confidence: 99%

Pathogenesis and Immune Response of Crimean-Congo Hemorrhagic Fever Virus in a STAT-1 Knockout Mouse Model

Bente

Alimonti

Shieh

et al. 2010

J Virol

210

242

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“…Although the first RT-PCR assays could identify viruses from most clades or geographic regions (Duh et al, 2006;Garrison et al, 2007;Wolfel et al, 2007), they were unable to recognize the highly divergent AP92 strain. However, a recently reported assay targeting a highly conserved portion of the 5 0 -noncoding region of the viral S segment, which is apparently required for ''panhandle'' formation and genome replication, is able to detect viruses in all 7 clades, including AP92 (Atkinson et al, 2012a).…”

Section: Diagnosismentioning

confidence: 99%

Crimean-Congo hemorrhagic fever: History, epidemiology, pathogenesis, clinical syndrome and genetic diversity

et al. 2013

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Bente, Dennis A.; Forrester, Naomi L.; Watts, Douglas M.; McAuley, Alexander J.; Whitehouse, Chris A.; and Bray, Mike, "CrimeanCongo hemorrhagic fever: History, epidemiology, pathogenesis, clinical syndrome and genetic diversity" (2013 b s t r a c tCrimean-Congo hemorrhagic fever (CCHF) is the most important tick-borne viral disease of humans, causing sporadic cases or outbreaks of severe illness across a huge geographic area, from western China to the Middle East and southeastern Europe and throughout most of Africa. CCHFV is maintained in vertical and horizontal transmission cycles involving ixodid ticks and a variety of wild and domestic vertebrates, which do not show signs of illness. The virus circulates in a number of tick genera, but Hyalomma ticks are the principal source of human infection, probably because both immature and adult forms actively seek hosts for the blood meals required at each stage of maturation. CCHF occurs most frequently among agricultural workers following the bite of an infected tick, and to a lesser extent among slaughterhouse workers exposed to the blood and tissues of infected livestock and medical personnel through contact with the body fluids of infected patients. CCHFV is the most genetically diverse of the arboviruses, with nucleotide sequence differences among isolates ranging from 20% for the viral S segment to 31% for the M segment. Viruses with diverse sequences can be found within the same geographic area, while closely related viruses have been isolated in far distant regions, suggesting that widespread dispersion of CCHFV has occurred at times in the past, possibly by ticks carried on migratory birds or through the international livestock trade. Reassortment among genome segments during co-infection of ticks or vertebrates appears to have played an important role in generating diversity, and represents a potential future source of novel viruses. In this article, we first review current knowledge of CCHFV, summarizing its molecular biology, maintenance and transmission, epidemiology and geographic range. We also include an extensive discussion of CCHFV genetic diversity, including maps of the range of the virus with superimposed phylogenetic trees. We then review the features of CCHF, including the clinical syndrome, diagnosis, treatment, pathogenesis, vaccine development and laboratory animal models of CCHF. The paper ends with a discussion of the possible future geographic range of the virus. For the benefit of researchers, we include a Supplementary Table listing all published reports of CCHF cases and outbreaks in the English-language literature, plus some principal articles in other languages, with total case numbers, case fatality rates and all CCHFV strains on GenBank.

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“…The extracted RNA was analyzed by CCHFV real-time RT-PCR, positive reactions were plotted against a standard curve, and the genome content/ml was calculated (24). The cycling reactions were performed in a Roche LightCycler 2.0 or 480 apparatus.…”

Section: Methodsmentioning

confidence: 99%

“…Samples were taken from patients displaying symptoms of CCHFV infection, and where possible, both acute-phase samples (A) and convalescentphase samples (B) were taken from the patients. All samples were also analyzed by conventional real-time RT-PCR as described by Wolfel et al (24).…”

Section: Methodsmentioning

confidence: 99%

Colorimetric Nucleic Acid Testing Assay for RNA Virus Detection Based on Circle-to-Circle Amplification of Padlock Probes

Zorzet

Göransson

et al. 2011

J Clin Microbiol

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We developed a molecular diagnostic method for detection of RNA virus based on padlock probes and colorimetric readout. The feasibility of our approach was demonstrated by using detection of Crimean-Congo hemorrhagic fever (CCHF) virus as a model. Compared with conventional PCR-based methods, our approach does not require advanced equipment, involves easier assay design, and has a sensitivity of 10 3 viral copies/ml. By using a cocktail of padlock probes, synthetic templates representing different viral strain variants could be detected. We analyzed 34 CCHF patient samples, and all patients were correctly diagnosed when the results were compared to those of the current real-time PCR method. This is the first time that highly specific padlock probes have been applied to detection of a highly variable target sequence typical of RNA viruses.

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Virus Detection and Monitoring of Viral Load in Crimean-Congo Hemorrhagic Fever Virus Patients

Cited by 116 publications

References 12 publications

Pathogenesis and Immune Response of Crimean-Congo Hemorrhagic Fever Virus in a STAT-1 Knockout Mouse Model

Pathogenesis and Immune Response of Crimean-Congo Hemorrhagic Fever Virus in a STAT-1 Knockout Mouse Model

Crimean-Congo hemorrhagic fever: History, epidemiology, pathogenesis, clinical syndrome and genetic diversity

Colorimetric Nucleic Acid Testing Assay for RNA Virus Detection Based on Circle-to-Circle Amplification of Padlock Probes

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