Immunoblot detection of antibodies to Toscana virus

Schwarz, Tino F.; Gilch, Sabine; Pauli, Christof; Jäger, Gundula

doi:10.1002/(sici)1096-9071(199606)49:2<83::aid-jmv2>3.0.co;2-f

Cited by 43 publications

(24 citation statements)

References 21 publications

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“…In fact, previous studies have revealed that the N protein of RVFV is a highly conserved among RVFV strains [19-21, 26, 27]. In addition, it was also reported that N protein is the main immunodominant viral protein in other members of the Bunyaviridae family [22,25]. Since rNp lacks infectivity with considerable stability, it is one of the suitable candidates for use as a diagnostic antigen in an ELISA [7,8,15,18].…”

Section: Discussionmentioning

confidence: 99%

Competitive ELISA for the Detection of Antibodies to Rift Valley Fever Virus in Goats and Cattle

Kim

Nah

Moon

et al. 2012

The Journal of Veterinary Medical Science

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ABSTRACT. Rift Valley fever virus (RVFV) is one of the important emerging viral diseases of serious impact in public health and animal hygiene both in human and animal industries. In this study, we developed a monoclonal antibody-based competitive ELISA for the detection of antibodies to RVFV in goats and cattle. The recombinant N protein of RVFV was expressed in E. coli with a six-histidine tag, and the purified N protein was used for detecting antigen with a competitive monoclonal antibody against RVFV antibodies. The competitive ELISA (C-ELISA) could detect antibodies at 9-11 days after inoculation in goats and cattle with a sensitivity of 94.7% (virus neutralization titer >32) and specificity of 99.7%, respectively. In addition, the C-ELISA did not show any cross-reactivity with positive sera against arboviruses such as Akabane, Aino, Chuzan, Ibaraki and bovine ephemeral fever virus, which are prevalent viral agents in ruminant animals throughout Southeast Asia. The results of the present study indicate that the C-ELISA is a simple, rapid and convenient serodiagnostic method for RVFV in goats and cattle.

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Section: Discussionmentioning

confidence: 99%

Competitive ELISA for the Detection of Antibodies to Rift Valley Fever Virus in Goats and Cattle

Kim

Nah

Moon

et al. 2012

The Journal of Veterinary Medical Science

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“…For this reason and the fact that the virus has recently been shown to spread beyond its traditional endemic areas (Shoemaker et al, 2002) there is an urgent need for the development of diagnostic methods allowing for safe and cost effective production and application of RVF immunoreagents. The N-protein of RVFV was chosen as an antigen because it was shown to be the immunodominant protein in other members of the Bunyaviridae family (Swanepoel et al, 1986b, Vapalahti et al, 1995and Schwarz et al, 1996.…”

Section: Discussionmentioning

confidence: 99%

Cloning and expression of Rift Valley fever virus nucleocapsid (N) protein and evaluation of a N-protein based indirect ELISA for the detection of specific IgG and IgM antibodies in domestic ruminants

Fafetine¹,

Tijhaar²,

Pawęska³

et al. 2007

Veterinary Microbiology

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“…The S segment encodes nucleocapsid (N) protein and a nonstructural (NS) protein using an ambisense coding strategy (7). Of all the genome-encoded proteins, N protein is the most immunodominant viral protein, and it is highly conserved in the Bunyaviridae family (9,15,17).…”

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confidence: 99%

Preparation and Evaluation of Recombinant Severe Fever with Thrombocytopenia Syndrome Virus Nucleocapsid Protein for Detection of Total Antibodies in Human and Animal Sera by Double-Antigen Sandwich Enzyme-Linked Immunosorbent Assay

et al. 2012

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The recent emergence of the human infection confirmed to be caused by severe fever with thrombocytopenia syndrome virus (SFTSV) in China is of global concern. Safe diagnostic immunoreagents for determination of human and animal seroprevalence in epidemiological investigations are urgently needed. This paper describes the cloning and expression of the nucleocapsid (N) protein of SFTSV. An N-protein-based double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) system was set up to detect the total antibodies in human and animal sera. We reasoned that as the double-antigen sandwich ELISA detected total antibodies with a higher sensitivity than traditional indirect ELISA, it could be used to detect SFTSV-specific antibodies from different animal species. The serum neutralization test was used to validate the performance of this ELISA system. All human and animal sera that tested positive in the neutralization test were also positive in the sandwich ELISA, and there was a high correlation between serum neutralizing titers and ELISA readings. Cross-reactivity was evaluated, and the system was found to be highly specific to SFTSV; all hantavirus-and dengue virus-confirmed patient samples were negative. SFTSV-confirmed human and animal sera from both Anhui and Hubei Provinces in China reacted with N protein in this ELISA, suggesting no major antigenic variation between geographically disparate virus isolates and the suitability of this assay in nationwide application. ELISA results showed that 3.6% of the human serum samples and 47.7% of the animal field serum samples were positive for SFTSV antibodies, indicating that SFTSV has circulated widely in China. This assay, which is simple to operate, poses no biohazard risk, does not require sophisticated equipment, and can be used in disease surveillance programs, particularly in the screening of large numbers of samples from various animal species.

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Immunoblot detection of antibodies to Toscana virus

Cited by 43 publications

References 21 publications

Competitive ELISA for the Detection of Antibodies to Rift Valley Fever Virus in Goats and Cattle

Competitive ELISA for the Detection of Antibodies to Rift Valley Fever Virus in Goats and Cattle

Cloning and expression of Rift Valley fever virus nucleocapsid (N) protein and evaluation of a N-protein based indirect ELISA for the detection of specific IgG and IgM antibodies in domestic ruminants

Preparation and Evaluation of Recombinant Severe Fever with Thrombocytopenia Syndrome Virus Nucleocapsid Protein for Detection of Total Antibodies in Human and Animal Sera by Double-Antigen Sandwich Enzyme-Linked Immunosorbent Assay

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