Determination of B cell epitopes and evaluation of antigen capture ELISA for the earlier diagnosis of CHIK virus using anti-rCHIK E1 rabbit antibodies
Autor(es): Yathi Krishna Kammara, Bhasker Salini, Chinnamma Mohankumar
Resumo: Chikungunya fever caused by an alpha virus has been generally considered as self limiting - non fatal. Recent reports on Chikungunya infection indicate high mortality rates due to the severity of the viral infection. For the early diagnosis of CHIK virus, the incubation period required for the development of antibodies in the serum of patients was a constraint for antigen based ELISA. The results of the present study demonstrates the development - evaluation of the antigen capture ELISA using recombinant anti-CHIK rabbit antibodies - anti-CHIK human antibody for more specific - rapid detection of CHIK viral antigen. A comprehensive bioinformatics analysis of the amino acid sequence of CHIK E1 protein was done for determining the antigenic residues, predominant B cell epitopes - their properties. Rabbit antibodies against recombinant CHIK E1 antigen was developed - purified. Antigen capture ELISA was done in 104 CHIK patient serum samples using anti-rCHIK E1 rabbit antibodies - anti-CHIK human antibodies. The highest rate of sensitivity (96%) - specificity (100%) was observed in the assay data - it highlights the accuracy of the test as a clinical diagnostic tool. No cross reactivity was observed with samples of dengue patients. Apart from the development - evaluation of the ELISA test, the dominant epitopes identified in the recombinant CHIK E1 protein sequence can be exploited for the development of a subunit Chikungunya vaccine.
Palavras-Chave: Chikungunya virus; E1 coat gene; Recombinant antigen; B cell epitopes; Anti-rCHIK E1 rabbit antibodies; Antigen capture ELISA
Imprenta: Journal of Immunological Methods, v. 393, n. 1-2, p. 45-52, 2013
Identificador do objeto digital: 10.1016/j.jim.2013.04.005
Descritores: Chikungunya virus - Cell ; Chikungunya virus - Molecular structure ; Chikungunya virus - Pathogenesis ; Chikungunya virus - Proteins ; Chikungunya virus - Antibodies ; Chikungunya virus - Viral infections ; Chikungunya Virus - Virus ; Chikungunya virus - Vaccine ; Chikungunya virus - Chikungunya fever ; Chikungunya virus - Dengue ; Chikungunya virus - Immunology ; Chikungunya virus - Public health
Data de publicação: 2013
