Early serodiagnosis of acute human Crimean-Congo hemorrhagic fever virus infections by novel capture assays

Emmerich, Petra; Avšič–Županc, Tatjana; Chinikar, Sadegh; Saksida, Ana; Thomé-Bolduan, Corinna; Parczany-Hartmann, Angela; Langroudi, Arash Ghalyanchi; Moradi, Maryam; Ahmeti, Salih; Günther, Stephan; Schmidt‐Chanasit, Jonas

doi:10.1016/j.jcv.2010.05.002

Cited by 12 publications

(9 citation statements)

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“…CCHFV NP has been used previously as an antigen in a variety of ELISA applications, either utilizing a combination of inactivated native virus lysate and an NP-specific monoclonal antibody [ 24 ] or purified recombinant NP produced either in insect cells [ 23 , 25 , 26 ], mammalian cells [ 27 ], E . coli [ 28 , 29 ] or even plants [ 30 ].…”

Section: Discussionmentioning

confidence: 99%

Sensitive and specific detection of Crimean-Congo Hemorrhagic Fever Virus (CCHFV)—Specific IgM and IgG antibodies in human sera using recombinant CCHFV nucleoprotein as antigen in μ-capture and IgG immune complex (IC) ELISA tests

et al. 2018

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As the most widespread tick-borne arbovirus causing infections in numerous countries in Asia, Africa and Europe, Crimean-Congo Hemorrhagic Fever Virus (CCHFV, family Nairoviridae) was included in the WHO priority list of emerging pathogens needing urgent Research & Development attention. To ensure preparedness for potential future outbreak scenarios, reliable diagnostic tools for identification of acute cases as well as for performance of seroprevalence studies are necessary. Here, the CCHFV ortholog of the major bunyavirus antigen, the nucleoprotein (NP), was recombinantly expressed in E.coli, purified and directly labeled with horseradish peroxidase (HRP). Employing this antigen, two serological tests, a μ-capture ELISA for the detection of CCHFV-specific IgM antibodies (BLACKBOX CCHFV IgM) and an IgG immune complex (IC) ELISA for the detection of CCHFV-specific IgG antibodies (BLACKBOX CCHFV IgG), were developed. Test performance was evaluated and compared with both in-house gold standard testing by IgM/IgG indirect immunofluorescence (IIF) and commercially available ELISA tests (VectoCrimean-CHF-IgM/IgG, Vector-Best, Russia) using a serum panel comprising paired samples collected in Kosovo during the years 2013–2016 from 15 patients with an acute, RT-PCR-confirmed CCHFV infection, and 12 follow-up sera of the same patients collected approximately one year after having overcome the infection. Reliably detecting IgM antibodies in all acute phase sera collected later than day 4 after onset of symptoms, both IgM ELISAs displayed excellent diagnostic and analytical sensitivity (100%, 95% confidence interval (CI): 85.2%–100.0%). While both IgG ELISAs readily detected the high IgG titers present in convalescent patients approximately one year after having overcome the infection (sensitivity 100%, 95% CI: 73.5%–100.0%), the newly developed BLACKBOX CCHFV IgG ELISA was superior to the commercial IgG ELISA in detecting the rising IgG titers during the acute phase of the disease. While all samples collected between day 11 and 19 after onset of symptoms tested positive in both the in-house gold standard IIFT and the BLACKBOX CCHFV IgG ELISA (sensitivity 100%, 95% CI: 71.5%–100.0%), only 27% (95% CI: 6.0%–61.0%) of those samples were tested positive in the commercial IgG ELISA. No false positive signals were observed in either IgM/IgG ELISA when analyzing a priori CCHFV IgM/IgG negative serum samples from healthy blood donors, malaria patients and flavivirus infected patients as well as CCHFV IgM/IgG IIFT negative serum samples from healthy Kosovar blood donors (for BLACKBOX CCHFV IgM/IgG: n = 218, 100% specificity, 95% CI: 98.3%–100.0%, for VectoCrimean-CHF-IgM/IgG: n = 113, 100% specificity, 95% CI: 96.8%–100.0%).

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Section: Discussionmentioning

confidence: 99%

Sensitive and specific detection of Crimean-Congo Hemorrhagic Fever Virus (CCHFV)—Specific IgM and IgG antibodies in human sera using recombinant CCHFV nucleoprotein as antigen in μ-capture and IgG immune complex (IC) ELISA tests

et al. 2018

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“…After 5 days of incubation, cells were fixed with 4% formaldehyde in phosphatebuffered saline (PBS), washed with water, and permeabilized with 0.5% Triton X-100 in PBS. After washing and blocking with 10% FCS in PBS, infected cell foci were detected with CCHFV nucleoprotein (NP)-specific monoclonal antibody A4 [48]. After washing, cells were incubated with peroxidase-labeled anti-mouse IgG.…”

Section: Titration Of Virus and Antibodiesmentioning

confidence: 99%

“…Cells were counterstained with hematoxylin. IHC with primary antibodies directed against CCHFV NP (monoclonal antibody A4 [48], 1:500) and neutrophil marker Ly6G (1:1,000; BD Bioscience) was performed manually. Sections were boiled in citrate buffer (pH 6) for 1 h and incubated with antibody at 4uC overnight.…”

Section: Histology and Immunohistochemistrymentioning

confidence: 99%

Evaluation of Antiviral Efficacy of Ribavirin, Arbidol, and T-705 (Favipiravir) in a Mouse Model for Crimean-Congo Hemorrhagic Fever

et al. 2014

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BackgroundMice lacking the type I interferon receptor (IFNAR−/− mice) reproduce relevant aspects of Crimean-Congo hemorrhagic fever (CCHF) in humans, including liver damage. We aimed at characterizing the liver pathology in CCHF virus-infected IFNAR−/− mice by immunohistochemistry and employed the model to evaluate the antiviral efficacy of ribavirin, arbidol, and T-705 against CCHF virus.Methodology/Principal FindingsCCHF virus-infected IFNAR−/− mice died 2–6 days post infection with elevated aminotransferase levels and high virus titers in blood and organs. Main pathological alteration was acute hepatitis with extensive bridging necrosis, reactive hepatocyte proliferation, and mild to moderate inflammatory response with monocyte/macrophage activation. Virus-infected and apoptotic hepatocytes clustered in the necrotic areas. Ribavirin, arbidol, and T-705 suppressed virus replication in vitro by ≥3 log units (IC50 0.6–2.8 µg/ml; IC90 1.2–4.7 µg/ml). Ribavirin [100 mg/(kg×d)] did not increase the survival rate of IFNAR−/− mice, but prolonged the time to death (p<0.001) and reduced the aminotransferase levels and the virus titers. Arbidol [150 mg/(kg×d)] had no efficacy in vivo. Animals treated with T-705 at 1 h [15, 30, and 300 mg/(kg×d)] or up to 2 days [300 mg/(kg×d)] post infection survived, showed no signs of disease, and had no virus in blood and organs. Co-administration of ribavirin and T-705 yielded beneficial rather than adverse effects.Conclusions/SignificanceActivated hepatic macrophages and monocyte-derived cells may play a role in the proinflammatory cytokine response in CCHF. Clustering of infected hepatocytes in necrotic areas without marked inflammation suggests viral cytopathic effects. T-705 is highly potent against CCHF virus in vitro and in vivo. Its in vivo efficacy exceeds that of the current standard drug for treatment of CCHF, ribavirin.

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“…The CCHFV N protein induces an early, strong and long-lasting immune response in humans and therefore the native or recombinant N protein is often used as the antigen in serological assays. Capture ELISAs for detection of CCHFV-specific IgM (μ-capture) and IgG (rheumatoid factor capture) have been developed 64 , while the recent development of an indirect ELISA for the parallel measurement of CCHFV IgG and IgM antibodies using recombinant N protein as antigen removes the reliance on polyclonal antibody preparations 65 .…”

Section: Advances In Clinical Virologymentioning

confidence: 99%

Recent advances in research on Crimean-Congo hemorrhagic fever

Papa

Mirazimi

Köksal

et al. 2015

Journal of Clinical Virology

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Crimean-Congo hemorrhagic fever (CCHF) is an expanding tick-borne hemorrhagic disease with increasing human and animal health impact. Immense knowledge was gained over the past 10 years mainly due to advances in molecular biology, but also driven by an increased global interest in CCHFV as an emerging/re-emerging zoonotic pathogen. In the present article we discuss the advances in research with focus on CCHF ecology, epidemiology, pathogenesis, diagnostics, prophylaxis and treatment. Despite tremendous achievements, future activities have to concentrate on the development of vaccines and antivirals/therapeutics to combat CCHF. Vector studies need to continue for better public and animal health preparedness and response. We conclude with a roadmap for future research priorities.

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Early serodiagnosis of acute human Crimean-Congo hemorrhagic fever virus infections by novel capture assays

Cited by 12 publications

References 10 publications

Sensitive and specific detection of Crimean-Congo Hemorrhagic Fever Virus (CCHFV)—Specific IgM and IgG antibodies in human sera using recombinant CCHFV nucleoprotein as antigen in μ-capture and IgG immune complex (IC) ELISA tests

Sensitive and specific detection of Crimean-Congo Hemorrhagic Fever Virus (CCHFV)—Specific IgM and IgG antibodies in human sera using recombinant CCHFV nucleoprotein as antigen in μ-capture and IgG immune complex (IC) ELISA tests

Evaluation of Antiviral Efficacy of Ribavirin, Arbidol, and T-705 (Favipiravir) in a Mouse Model for Crimean-Congo Hemorrhagic Fever

Recent advances in research on Crimean-Congo hemorrhagic fever

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