1. Cloning, expression & evaluation of potential immunogenic recombinant capsid premembrane protein of West Nile virus
- Author
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Divanyshi Karothia, Sivakumar R. Rathinam, Manmohan Parida, and Jyoti S. Kumar
- Subjects
0301 basic medicine ,viruses ,030106 microbiology ,lcsh:Medicine ,Enzyme-Linked Immunosorbent Assay ,Biology ,General Biochemistry, Genetics and Molecular Biology ,law.invention ,Dengue fever ,03 medical and health sciences ,0302 clinical medicine ,Plaque reduction neutralization test ,Viral Envelope Proteins ,law ,medicine ,blood samples ,Humans ,CprM gene - blood samples - ELISA - plaque reduction neutralization test - recombinant protein - West Nile virus ,Serologic Tests ,030212 general & internal medicine ,Vector (molecular biology) ,CprM gene ,Cloning, Molecular ,Viral encephalitis ,lcsh:R ,virus diseases ,General Medicine ,plaque reduction neutralization test ,medicine.disease ,biology.organism_classification ,Virology ,Recombinant Proteins ,Flavivirus ,Capsid ,Immunoglobulin M ,Antibody Formation ,Recombinant DNA ,biology.protein ,Original Article ,ELISA ,Capsid Proteins ,West Nile virus ,recombinant protein ,West Nile Fever - Abstract
Background & objectives: West Nile virus (WNV) is a neurotropic flavivirus that has emerged globally as a significant cause of viral encephalitis. The early confirmatory diagnosis of WNV infections is important for timely clinical management and in areas where multiple flaviviruses are endemic. Diagnosis of WNV infection is primarily based on serodiagnosis, followed by virus isolation and identification. The aim of this study was to develop and evaluate a highly sensitive and specific immunoglobulin M (IgM) ELISA using the recombinant CprM protein (rWNV-CprM) for rapid, early and accurate diagnosis of WNV. Methods: The gene coding for the CprM protein of WNV was cloned and expressed in pET 28a vector followed by purification. An indirect IgM microplate ELISA using purified rWNV-CprM protein was optimized having no cross-reactivity with healthy human serum and serum samples obtained from patients with dengue and Japanese encephalitis viruses infection. Results: The comparative evaluation of this rWNV-CprM protein-specific IgM ELISA with plaque reduction neutralization test using 105 blood samples collected from patients suspected to have acute WNV infection revealed 98 per cent concordance with sensitivity and specificity of 100 and 97 per cent, respectively. Interpretation & conclusions: The recombinant CprM protein-based WNV-specific ELISA reported in this study may be useful for rapid screening of large numbers of blood samples in endemic areas during outbreaks.
- Published
- 2019