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Development of a enterovirus diagnostic assay system for diagnosis of viral myocarditis in humans
Byung-Kwan Lim1, Eun-Seon Ju, Dieu Hung Lao
1Division of Cardiology, Samsung Medical Center, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Seoul, Korea.
Insights
A new diagnostic system uses specific coxsackievirus B3 (CVB3) peptide sequences to detect CVB3 infection. This fast and accurate ELISA method aids in diagnosing viral myocarditis and dilated cardiomyopathy caused by enteroviruses.
Area of Science:
- Virology
- Immunology
- Biotechnology
Background:
- Coxsackievirus B3 (CVB3) is a major cause of chronic myocarditis and dilated cardiomyopathy.
- Current diagnostic methods for CVB3 infection lack effectiveness in humans.
Purpose of the Study:
- To establish a fast and accurate diagnostic system for CVB3 infection using capsid-protein-specific peptides.
- To develop an enzyme-linked immunosorbent assay (ELISA) for detecting CVB3 in patient sera.
Main Methods:
- Computationally predicted and synthesized peptide sequences from CVB3 capsid proteins (VP1 and VP2).
- Induction of immunoglobulin G (IgG) antibodies in rabbits immunized with synthesized peptides.
- Application of peptides in an ELISA system to detect anti-CVB3 antibodies in mouse and human serum samples.
Main Results:
- Synthesized peptides (VP1 and VP2) successfully induced specific antibody production in rabbits.
- The developed ELISA system accurately detected anti-CVB3 IgG antibodies in experimentally infected mouse serum.
- A VP2 peptide-based ELISA identified CVB3 infection in patients with confirmed CVB3-induced fulminant myocarditis.
Conclusions:
- The novel peptide sequences specifically interact with anti-CVB3 IgG antibodies.
- The developed ELISA system shows promise for the clinical diagnosis of enterovirus-induced myocarditis.
Abstract:
The coxsackieviruses type B3 (CVB3) are members of the genus Enterovirus of the family Picornaviridae. They are the commonest cause of chronic myocarditis and dilated cardiomyopathy. However, there is still no effective method for diagnosing CVB3 infection in humans. Here, a fast and accurate system that uses a capsid-protein-specific peptide sequence to detect CVB3 in the sera of patients with viral myocarditis was established. The peptide sequence was selected from the whole CVB3 capsid protein sequence by computationally predicting fragments with high antigenicity and low hydrophobicity. Two of eight possible peptide sequences were selected and commercially synthesized. The synthesized peptides encoded either the VP2 or VP1 capsid protein and induced immunoglobulin G antibody expression in immunized rabbits. Anti-VP2 and anti-VP1 sera detected the viral proteins extracted from CVB3-infected HeLa cells. The newly synthesized peptides successfully induced antibody production. These peptides, applied in an ELISA system, detected anti-CVB3 antibodies in virus-infected mouse serum. Moreover, an ELISA system based on the VP2 peptide detected CVB3 infection in patients with positively identified CVB3-induced fulminant myocarditis. These results indicate that these new peptides specifically interact with anti-CVB3 IgG antibodies in mouse and human sera. This ELISA system should be useful for the clinical diagnosis of enterovirus-induced myocarditis.
