title: Cloning, expression and evaluation of diagnostic potential of recombinant capsid protein based IgM ELISA for chikungunya virus.
abstract: The resurgence of chikungunya virus in the form of unprecedented explosive epidemic with unusual clinical severity after a gap of 32 years is a point of major public health concern. Definitive diagnosis is critical in differentiating the disease, especially in dengue endemic areas. The immunoglobulin M (IgM) enzyme-linked immunosorbent assay (ELISA) is widely used for diagnosis of chikungunya infection. However IgM ELISA based on whole virus antigen is associated with biohazard risk. The present study describes the development and evaluation of recombinant capsid protein based indirect IgM antibody capture micro plate enzyme linked immunosorbent assay (ELISA) for rapid and accurate diagnosis of chikungunya infection. The gene coding for capsid protein was cloned in frame with GST tag in pET41a+ vector and expressed in E. coli followed by purification with affinity chromatography. The comparative evaluation of in-house chikungunya IgM ELISA vis-a-vis commercially available SD ELISA kit with 90 chikungunya suspected acute phase human patient serum samples revealed 97% accordance. The overall sensitivity and specificity of the reported capsid protein based IgM ELISA was 100% and 95% respectively with 96% PPV and 100% NPV. These findings clearly demonstrated the usefulness of the recombinant capsid protein based CHIKV IgM ELISA for reliable clinical diagnosis of CHIKV infection in human patient.
authors.affiliation: Division of Virology, Defence Research & Development Establishment, Gwalior 474002, India.
authors.forename: Raj
authors.initials: R
authors.lastname: Priya
authors.affiliation: Division of Virology, Defence Research & Development Establishment, Gwalior 474002, India.
authors.forename: Mohsin
authors.initials: M
authors.lastname: Khan
authors.affiliation: Biochemistry Division, Defence Research & Development Establishment, Gwalior 474002, India.
authors.forename: M Kameswara
authors.initials: MK
authors.lastname: Rao
authors.affiliation: Division of Virology, Defence Research & Development Establishment, Gwalior 474002, India. Electronic address: paridamm@rediffmail.com.
authors.forename: Manmohan
authors.initials: M
authors.lastname: Parida
mesh_terms: D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000235:genetics; D036022:Capsid Proteins / Q000235:genetics; D065632:Chikungunya Fever / Q000175:diagnosis* / Q000276:immunology; D002646:Chikungunya virus / Q000235:genetics / Q000276:immunology*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D007075:Immunoglobulin M; D011994:Recombinant Proteins
keywords: Chikungunya; IgM ELISA; Recombinant capsid protein