Recombinant Nucleoprotein-Based Enzyme-Linked Immunosorbent Assay for Detection of Immunoglobulin G Antibodies to Crimean-Congo Hemorrhagic Fever Virus

Saijo, Masayuki; Tang, Qing; Niikura, Masahiro; Maeda, Akihiko; Ikegami, Tetsuro; Préhaud, Christophe; Kurane, Ichiro; Morikawa, Shigeru

doi:10.1128/jcm.40.5.1587-1591.2002

Cited by 81 publications

(97 citation statements)

References 20 publications

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“…The cDNAs of the SFTSV Gn/Gc protein were obtained from SFTSV (HB29 strain) by reverse transcription-PCR (RT-PCR). The Gn/Gc cDNA was cloned into the expression vector pKS336 (14). The resulting plasmid was designated pKS-SFTSV-GP.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Glycoprotein-Mediated Entry of Severe Fever with Thrombocytopenia Syndrome Virus

Tani

Shimojima

Fukushi

et al. 2016

J Virol

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Severe fever with thrombocytopenia syndrome (SFTS) is an emerging hemorrhagic fever with a high case fatality rate caused by SFTS virus (SFTSV). Effective vaccines and specific therapies for SFTS are urgently sought, and investigation into virus-host cell interactions is expected to contribute to the development of antiviral strategies. In this study, we have developed a pseudotype vesicular stomatitis virus (VSV) bearing the unmodified Gn/Gc glycoproteins (GPs) of SFTSV (SFTSVpv). We have analyzed the host cell entry of this pseudotype virus and native SFTSV. Both SFTSVpv and SFTSV exhibited high infectivity in various mammalian cell lines. The use of lysosomotropic agents indicated that virus entry occurred via pH-dependent endocytosis. SFTSVpv and SFTSV infectivity was neutralized by serial dilutions of convalescent-phase patient sera. Entry of SFTSVpv and growth of SFTSV were increased in Raji cells expressing not only the C-type lectin dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) but also DC-SIGN-related (DC-SIGNR) and liver and lymph node sinusoidal endothelial cell C-type lectin (LSECtin). 25-Hydroxycholesterol (25HC), a soluble oxysterol metabolite, inhibited the cell entry of SFTSVpv and the membrane fusion of SFTSV. These results indicate that pH-dependent endocytosis of SFTSVpv and SFTSV is enhanced by attachment to certain C-type lectins. SFTSVpv is an appropriate model for the investigation of SFTSV-GP-mediated cell entry and virus neutralization at lower biosafety levels. Furthermore, 25HC may represent a potential antiviral agent against SFTS. IMPORTANCESFTSV is a recently discovered bunyavirus associated with SFTS, a viral hemorrhagic fever with a high case fatality rate endemic to China, South Korea, and Japan. Because little is known about the characteristics of the envelope protein and entry mechanisms of SFTSV, further studies will be required for the development of a vaccine or effective therapies. In this study, we investigated the mechanism of SFTSV cell entry using SFTSVpv and the native virus. SFTSV can grow in nonsusceptible cell lines in the presence of certain C-type lectins. Moreover, 25HC, an oxysterol metabolite, may represent a potential therapeutic inhibitor of SFTSV infection. Severe fever with thrombocytopenia syndrome (SFTS), a newly recognized emerging viral infectious disease, is caused by a novel phlebovirus in the family Bunyaviridae, SFTS virus (SFTSV) (1-3). SFTSV can be transmitted to humans by tick bites, causing abrupt high fever, thrombocytopenia, leukopenia, and gastrointestinal symptoms, with a high case fatality rate (4). SFTSV is a single-stranded negative-sense RNA virus with three genomic segments, L, M, and S. The L segment encodes a viral RNA polymerase, while the M segment encodes the two viral envelope glycoproteins (GPs) Gn and Gc. The S segment is an ambisense RNA encoding a nucleoprotein (NP) and a nonstructural protein (NSs). The Gn and Gc proteins of bunyaviruses are usually localized in the Golgi app...

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Section: Methodsmentioning

confidence: 99%

Characterization of Glycoprotein-Mediated Entry of Severe Fever with Thrombocytopenia Syndrome Virus

Tani

Shimojima

Fukushi

et al. 2016

J Virol

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“…An antibody response is rarely detectable in fatal cases and diagnosis is usually confirmed by isolation of the virus from the serum or liver biopsy specimens. Recently, new immunological assays incorporating recombinant CCHFV nucleoprotein have been developed and used in an IFA (Saijo et al, 2002b) or in an ELISA (Saijo et al, 2002a;Tang et al, 2003) to detect serum antibodies from infected patients.…”

Section: Immunological Assaysmentioning

confidence: 99%

Crimean–Congo hemorrhagic fever

2004

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Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne disease caused by the arbovirus Crimean-Congo hemorrhagic fever virus (CCHFV), which is a member of the Nairovirus genus (family Bunyaviridae). CCHF was first recognized during a large outbreak among agricultural workers in the mid-1940s in the Crimean peninsula. The disease now occurs sporadically throughout much of Africa, Asia, and Europe and results in an approximately 30% fatality rate. After a short incubation period, CCHF is characterized by a sudden onset of high fever, chills, severe headache, dizziness, back, and abdominal pains. Additional symptoms can include nausea, vomiting, diarrhea, neuropsychiatric, and cardiovascular changes. In severe cases, hemorrhagic manifestations, ranging from petechiae to large areas of ecchymosis, develop. Numerous genera of ixodid ticks serve both as vector and reservoir for CCHFV; however, ticks in the genus Hyalomma are particularly important to the ecology of this virus. In fact, occurrence of CCHF closely approximates the known world distribution of Hyalomma spp. ticks. Therefore, exposure to these ticks represents a major risk factor for contracting disease; however, other important risk factors are known and are discussed in this review. In recent years, major advances in the molecular detection of CCHFV, particularly the use of real-time reverse transcription-polymerase chain reaction (RT-PCR), in clinical and tick samples have allowed for both rapid diagnosis of disease and molecular epidemiology studies. Treatment options for CCHF are limited. Immunotherapy and ribavirin have been tried with varying degrees of success during sporadic outbreaks of disease, but no case-controlled trials have been conducted. Consequently, there is currently no antiviral treatment for CCHF approved by the U.S. Food and Drug Administration (FDA). However, renewed interested in CCHFV, as well as increased knowledge of its basic biology, may lead to improved therapies in the future. This article reviews the history, epidemiology, ecology, clinical features, pathogenesis, diagnosis, and treatment of CCHF. In addition, recent advances in the molecular biology of CCHFV are presented, and issues related to its possible use as a bioterrorism agent are discussed.

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“…ELISA metodları spesifik ve hassastır (17). Son zamanlarda, rekombinant-nükleoprotein bazlı IgG ELISA kiti, KKKAV enfeksiyonlarının teşhisinde kullanım amaçlı geliştirilmiştir (18,19 (21). Ayrıca doğal antijen olarak kullanılmaları açısından özgül ve hassastırlar (18,22).…”

Section: Discussionunclassified

Kırım Kongo kanamalı ateşi virüsü nükleoproteininin rekombinant olarak elde edilmesi ve ELISA'da kullanımı

Dönmez¹,

Pınarbaşı²

2016

Ege Tıp Dergisi

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Recombinant Nucleoprotein-Based Enzyme-Linked Immunosorbent Assay for Detection of Immunoglobulin G Antibodies to Crimean-Congo Hemorrhagic Fever Virus

Cited by 81 publications

References 20 publications

Characterization of Glycoprotein-Mediated Entry of Severe Fever with Thrombocytopenia Syndrome Virus

Characterization of Glycoprotein-Mediated Entry of Severe Fever with Thrombocytopenia Syndrome Virus

Crimean–Congo hemorrhagic fever

Kırım Kongo kanamalı ateşi virüsü nükleoproteininin rekombinant olarak elde edilmesi ve ELISA'da kullanımı

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