Detection of Crimean–Congo Hemorrhagic Fever, Hanta, and Sandfly Fever Viruses by Real-Time RT-PCR

Ibrahim, Sofi; Aitichou, Mohamed; Hardick, Justin; Blow, Jamie A.; O’Guinn, Monica L.; Schmaljohn, Connie S.

doi:10.1007/978-1-60761-817-1_19

Cited by 13 publications

(7 citation statements)

References 17 publications

(3 reference statements)

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“…Direct diagnosis of CCHFV infection is based either on virus isolation or on molecular detection of the genome (41)(42)(43)(44)(45). However, the period in which the virus can be detected in patients is rather short.…”

Section: Generation Of Cchf Virus-like Particles Expressing a Reportementioning

confidence: 99%

A Virus-Like Particle System Identifies the Endonuclease Domain of Crimean-Congo Hemorrhagic Fever Virus

Devignot

Bergeron

Nichol

et al. 2015

J Virol

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Crimean-Congo hemorrhagic fever virus (CCHFV; genus Nairovirus) is an extremely pathogenic member of the Bunyaviridae family. Since handling of the virus requires a biosafety level 4 (BSL-4) facility, little is known about pathomechanisms and host interactions. Here, we describe the establishment of a transcriptionally competent virus-like particle (tc- VLP IMPORTANCECrimean-Congo hemorrhagic fever virus (CCHFV) is an extremely virulent pathogen of humans. Since the virus can be handled only at the highest biosafety level, research is restricted to a few specialized laboratories. We developed a plasmid-based system to produce virus-like particles with the ability to infect cells and transcribe a reporter genome. Due to the absence of viral genes, the virus-like particles are unable to spread or cause disease, thus allowing study of aspects of CCHFV biology under relaxed biosafety conditions. C rimean-Congo hemorrhagic fever (CCHF) is a severe viral disease in Eastern Europe, the Middle East, Asia, and Africa, reported to have a case/fatality rate of approximately 30% (1). Infections of humans are associated with an acute febrile disease that can lead to hemorrhages, hypovolemic shock, and death, while in infected animals no clinical signs can be detected. The CCHF virus (CCHFV) is transmitted via tick bites (principally from the Hyalomma genus) or by direct contact with blood or tissues from infected persons or animals (2). The efficiency of ribavirin as an antiviral in humans is still under debate (3), and there are currently no established prophylaxis, no specific treatment, and no FDA-approved vaccine. The pathogenesis as well as immune responses are poorly characterized, mostly due to the restriction to high-biosafety-level facilities (biosafety level 4 [BSL-4]) for handling of virus and biological samples. The only animal models available so far are based on mice lacking antiviral interferon (IFN) responses, thus hampering studies on innate immune system interactions (4, 5). Clearly, there is paucity in tools and methods to better study the virus and its host cell interactions.CCHFV belongs to the Nairovirus genus of the family Bunyaviridae. This enveloped virus has a single-stranded negative-sense RNA genome (viral RNA [vRNA]) of an unusually large total size of 19,146 nucleotides (nt). The genome is divided into 3 segments and encodes 4 structural proteins: the RNA-dependent RNA polymerase (RdRp) L on the large (L) segment (12,108 nt), the two glycoproteins (GPs) Gc and Gn on the medium (M) segment (5,366 nt), and the nucleoprotein N on the small (S) segment (1,672 nt). The nucleoprotein N binds each RNA segment to form ribonucleoprotein complexes (RNPs), the functional template of the viral polymerase (pol) L for transcription and replication. Shortly after infection, CCHFV performs the so-called primary transcription leading to initial mRNA synthesis. The protein products are necessary for subsequent replication of the viral genome, resulting in copy RNA (cRNA) (positive sense) and progeny genom...

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Section: Generation Of Cchf Virus-like Particles Expressing a Reportementioning

confidence: 99%

A Virus-Like Particle System Identifies the Endonuclease Domain of Crimean-Congo Hemorrhagic Fever Virus

Devignot

Bergeron

Nichol

et al. 2015

J Virol

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“…However, because of the remarkable genetic variability among CCHFV strains, most current RT-PCRs either fail to detect specific strains or lack sensitivity [18]. A further improvement has been the development of real-time RT-PCRs, which have higher specificity and sensitivity, higher time-effectiveness and lower risk of contamination than conventional RT-PCR [19]–[21].…”

Section: Introductionmentioning

confidence: 99%

First International External Quality Assessment of Molecular Detection of Crimean-Congo Hemorrhagic Fever Virus

et al. 2012

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Crimean-Congo hemorrhagic fever (CCHF) is a zoonosis caused by a Nairovirus of the family Bunyaviridae . Infection is transmitted to humans mostly by Hyalomma ticks and also by direct contact with the blood or tissues of infected humans or viremic livestock. Clinical features usually include a rapid progression characterized by hemorrhage, myalgia and fever, with a lethality rate up to 30%. CCHF is one of the most widely distributed viral hemorrhagic fevers and has been reported in Africa, the Middle East and Asia, as well as parts of Europe. There is no approved vaccine or specific treatment against CCHF virus (CCHFV) infections. In this context, an accurate diagnosis as well as a reliable surveillance of CCHFV infections is essential. Diagnostic techniques include virus culture, serology and molecular methods, which are now increasingly used. The European Network for the Diagnostics of “Imported” Viral Diseases organized the first international external quality assessment of CCHVF molecular diagnostics in 2011 to assess the efficiency and accurateness of CCHFV molecular methods applied by expert laboratories. A proficiency test panel of 15 samples was distributed to the participants including 10 different CCHFV preparations generated from infected cell cultures, a preparation of plasmid cloned with the nucleoprotein of CCHFV, two CCHFV RNA preparations and two negative controls. Forty-four laboratories worldwide participated in the EQA study and 53 data sets were received. Twenty data sets (38%) met all criteria with optimal performance, 10 (19%) with acceptable performance, while 23 (43%) reported results showing a need for improvement. Differences in performance depended on the method used, the type of strain tested, the concentration of the sample tested and the laboratory performing the test. These results indicate that there is still a need for improving testing conditions and standardizing protocols for the molecular detection of Crimean-Congo hemorrhagic fever virus.

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“…Others have previously detected hantavirus by RT-qPCR but to our knowledge, this is the first effort to validate this technique as a diagnostic tool in a large number of patients (Evander et al 2007; Ibrahim et al 2011; Machado and Souza 2013). …”

Section: Discussionmentioning

confidence: 98%

Molecular method for the detection of Andes hantavirus infection: validation for clinical diagnostics

Vial

Martínez-Valdebenito

Rios

et al. 2016

Diagnostic Microbiology and Infectious Disease

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Hantavirus Cardiopulmonary Syndrome is a severe disease caused by exposure to New World hantaviruses. Early diagnosis is difficult due to the lack of specific initial symptoms. Anti-hantavirus antibodies are usually negative until late in the febrile prodrome or the beginning of cardiopulmonary phase while Andes hantavirus (ANDV) RNA genome can be detected before symptoms onset. We analyzed the effectiveness of RTqPCR as a diagnostic tool detecting ANDV-Sout genome in peripheral blood cells from 78 confirmed hantavirus patients and 166 negative controls. Our results indicate that RTqPCR had a low detection limit (~10 copies), with a specificity of 100% and a sensitivity of 94.9%. This suggests the potential for establishing RT-qPCR as the assay of choice for early diagnosis, promoting early effective care of patients and improve other important aspects of ANDV infection management, such as compliance of biosafety recommendations for health personnel in order to avoid nosocomial transmission.

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Detection of Crimean–Congo Hemorrhagic Fever, Hanta, and Sandfly Fever Viruses by Real-Time RT-PCR

Cited by 13 publications

References 17 publications

A Virus-Like Particle System Identifies the Endonuclease Domain of Crimean-Congo Hemorrhagic Fever Virus

A Virus-Like Particle System Identifies the Endonuclease Domain of Crimean-Congo Hemorrhagic Fever Virus

First International External Quality Assessment of Molecular Detection of Crimean-Congo Hemorrhagic Fever Virus

Molecular method for the detection of Andes hantavirus infection: validation for clinical diagnostics

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