Expression of the Lassa Virus Nucleocapsid Protein in Insect Cells Infected With a Recombinant Baculovirus: Application to Diagnostic Assays for Lassa Virus Infection

Barber, Glen N.; Clegg, J. C. S.; Lloyd, Graham

doi:10.1099/0022-1317-71-1-19

Cited by 35 publications

(16 citation statements)

References 31 publications

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“…Several recombinant NPs of haemorrhagic fever viruses have been produced by using bacterial or baculovirus expression systems (Barber et al, 1990 ;Go$ tt et al, 1991 ;KallioKokko et al, 1993 ;Zo$ ller et al, 1993 a, b). These recombinants were efficiently recognized by patient sera, but had a tendency to form aggregates or to be insoluble, which jeopardizes the reproducibility of results with batches.…”

Section: Discussionmentioning

confidence: 99%

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Recombinant Ebola virus nucleoprotein and glycoprotein (Gabon 94 strain) provide new tools for the detection of human infections.

Préhaud

Hellebrand

Coudrier

et al. 1998

Journal of General Virology

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After cloning and sequencing the glycoprotein (GP) gene of one of the Gabonese strains of Ebola virus isolated during the 1994-1996 outbreak, it was shown that the circulating virus was of the Zaire subtype. This was confirmed in this study by cloning and sequencing the nucleoprotein (NP) gene of this strain. These two structural proteins were also expressed as recombinant proteins and used in ELISA tests. NP was expressed as a His-tagged fusion protein in Escherichia coli and was purified on resins charged with nickel ions. GP was expressed

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Section: Discussionmentioning

confidence: 99%

“…In other forms of haemorrhagic fevers, i.e. those due to hantaviruses and to the arenavirus Lassa virus, the NP has been identified as a major antigen, inducing an early and long-lasting immune response (Barber et al, 1990 ;Lloyd et al, 1989 ;Zo$ ller et al, 1993 a, b).…”

Section: Introductionmentioning

confidence: 99%

Recombinant Ebola virus nucleoprotein and glycoprotein (Gabon 94 strain) provide new tools for the detection of human infections.

Préhaud

Hellebrand

Coudrier

et al. 1998

Journal of General Virology

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“…This may be explained by the high serum dilutions of about 1:≥100, which have to be applied to avoid non-specific background staining. Moreover, in the case of recombinant proteins (Barber et al, 1990;Gunther et al, 2001;ter Meulen et al, 1998) the loss of virus-specific epitopes may also contribute to a reduced sensitivity.…”

Section: Introductionmentioning

confidence: 96%

“…As an alternative to IFA, indirect ELISA techniques have been published to detect IgG and/or IgM antibodies using tissue culture (Bausch et al, 2000;Jahrling et al, 1985b;Niklasson et al, 1984) or by recombinant antigens (Barber et al, 1990;Gunther et al, 2001;ter Meulen et al, 1998). Anti-Lassa IgG antibodies were detected by indirect ELISAs.…”

Section: Introductionmentioning

confidence: 98%

Reverse ELISA for IgG and IgM antibodies to detect Lassa virus infections in Africa

Emmerich

Thomé-Bolduan

Drosten

et al. 2006

Journal of Clinical Virology

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“…Serosurveys reveal a prevalence of Lassa antibodies of up to 50% in some areas of West Africa [7,8,10,13]. Lassa virus expresses only four proteins, two of which ± nucleoprotein (NP) and glycoprotein ± have been shown to induce antibodies following infection [2,6,7,10]. An NP-speci®c T cell response has recently been demonstrated in persons from Lassa fever endemic regions with serological evidence of previous Lassa virus infection [11].…”

Section: Introductionmentioning

confidence: 99%

Antibodies to Lassa virus Z protein and nucleoprotein co-occur in human sera from Lassa fever endemic regions

Günther

Kühle

Rehder

et al. 2001

Med Microbiol Immunol

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It is not known whether the small 11-kDa Z protein of Lassa virus is immunogenic during human Lassa virus infection. To obtain evidence for the existence of an antibody response and to test the suitability of these antibodies for serosurveys, sera from Lassa fever endemic regions (Guinea and Nigeria, n = 75) were tested for co-reactivity to Z protein and nucleoprotein (NP). Sera from a non-endemic region (Uganda, n = 50) served as a specificity control. Z protein and NP were expressed in Escherichia coli, affinity-purified, and used as antigen in Western blot. Indirect immunofluorescence (IIF) with Lassa virus-infected cells was performed for comparison. Due to high unspecific reactivity of the African sera, Western blot testing was performed with a 1:1,000 serum dilution. Under these conditions, none of the control sera but 12% of the sera from endemic regions co-reacted with both Z protein and NP. Reactivity to Z protein was significantly associated with NP reactivity (P < 10(-6)). NP and Z protein-specific antibodies were co-detected in 33% of the IIF-positive sera and in 5% of the IIF-negative sera (P = 0.001). These data provide evidence for appearance of antibodies to Z protein and NP following Lassa virus infection. A recombinant blot for detection of both antibody specificities seems to be specific but less sensitive than IIF.

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Expression of the Lassa Virus Nucleocapsid Protein in Insect Cells Infected With a Recombinant Baculovirus: Application to Diagnostic Assays for Lassa Virus Infection

Cited by 35 publications

References 31 publications

Recombinant Ebola virus nucleoprotein and glycoprotein (Gabon 94 strain) provide new tools for the detection of human infections.

Recombinant Ebola virus nucleoprotein and glycoprotein (Gabon 94 strain) provide new tools for the detection of human infections.

Reverse ELISA for IgG and IgM antibodies to detect Lassa virus infections in Africa

Antibodies to Lassa virus Z protein and nucleoprotein co-occur in human sera from Lassa fever endemic regions

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