Yonsei Med J.  2008 Oct;49(5):828-835. 10.3349/ymj.2008.49.5.828.

Expression and Evaluation of Chikungunya Virus E1 and E2 Envelope Proteins for Serodiagnosis of Chikungunya Virus Infection

• Cho B
• Jeon BY
• Kim J
• Noh J
• Kim J
• Park M
• Park S

• KMID: 2158202
• DOI: http://doi.org/10.3349/ymj.2008.49.5.828

Abstract

PURPOSE
Chikungunya virus (CHIKV) causes endemic or epidemic outbreaks of CHIKV fever, which is a mosquitoe-transmitted viral disease in Africa, India, South-East Asia, and recently Southern Europe. Currently, serological diagnostic tests such as hemagglutination inhibition test (HI test), in-house IgM capture enzyme-linked immunosorbent assays (ELISA), and indirect immunofluorescence test were used for diagnosis of chikungunya fever, which are based on whole virus antigens. MATERIALS AND METHODS: CHIKV E1, and E2 envelope proteins for the CHIKV-specific serodiagnostic reagents for chikungunya fever were expressed in baculovirus expression system. The seroreactivity of recombinant CHIKV E1 and E2 envelope proteins were evaluated using sera panels of patients from Laboratoire Marcel Merieux by indirect IgM capture ELISA. RESULTS: The recombinant CHIKV E1 and E2 envelope protein showed sensitivity of 77.5% and 90%, respectively. The specificities of both CHIKV E1 and E2 envelope proteins were 100%. CONCLUSION: The recombinant CHIKV E1 and E2 envelope proteins could be a useful diagnostic reagent for CHIKV infection.

Keyword

Chikungunya virus; E1 and E2; envelope protein; baculovirus; indirect IgM ELISA

MeSH Terms

Alphavirus Infections/*diagnosis
Animals
Baculoviridae/genetics/metabolism
Cells, Cultured
Chikungunya virus/genetics
Cloning, Molecular
Enzyme-Linked Immunosorbent Assay/methods
Recombinant Proteins/immunology
Sensitivity and Specificity
Serologic Tests/*methods
Viral Envelope Proteins/*immunology

Figure

• 

  Fig. 1 Analysis of expression of CHIKV E1 (A) and E2 (B) envelope protein in Sf9 cells. Sf9 cell lysates were analysed by SDS-PAGE, and the gel was stained with Coomassie blue (lanes 1 to 3) or analysed by Western blot with pooled anti-CHIKV positive serum (lane 4). M; Prestained protein marker, lane 1; lysate of Sf9 cells infected with mock-vector, lane 2; lysate of Sf9 cells infected with recombinant virus containing CHIKV E1 (A) or E2 (B) envelope protein gene, lane 3; purified recombinant CHIKV E1 (A) or E2 (B) envelope protein, lane 4; Western blot analysis with pooled anti-CHIKV positive serum. CHIKV, chikungunya virus.

• 

  Fig. 2 Detection of anti-CHIKV IgM antibodies by recombinant CHIKV E1 (A) and E2 (B) envelope protein-based ELISA. Each well of a microtiter plate was coated with various amounts of recombinant CHIKV E1 and E2 envelope protein and evaluated for their reactivity with pooled anti-CHIKV positive or negative serum. Absorbance was read at 450 nm. The P/N ratio is A450 of positive serum/A450 of negative serum. CHIKV, chikungunya virus.

• 

  Fig. 3 The seroreactivity of the recombinant CHIKV E1 and E2 envelope proteins using indirect IgM ELISA. Forty anti-CHIKV positive serum samples were used to evaluate the seroreactivity of the recombinant CHIKV E1 (•) and E2 (•) envelope proteins. The absorbance was read at 450 nm. IgM capture ELISA data (○) were supplied from Laboratoire Marcel Merieux. CHIKV, chikungunya virus.

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