Recombinant hepatitis A virus antigen: improved production and utility in diagnostic immunoassays

F D LaBrecque; D R LaBrecque; D Klinzman; S Perlman; J B Cederna; P L Winokur; J Q Han; J T Stapleton

doi:10.1128/JCM.36.7.2014-2018.1998

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Recombinant hepatitis A virus antigen: improved production and utility in diagnostic immunoassays

Journal article

Open access

Peer reviewed

Recombinant hepatitis A virus antigen: improved production and utility in diagnostic immunoassays

F D LaBrecque, D R LaBrecque, D Klinzman, S Perlman, J B Cederna, P L Winokur, J Q Han and J T Stapleton

Journal of clinical microbiology, Vol.36(7), pp.2014-2018

07/1998

DOI: 10.1128/JCM.36.7.2014-2018.1998

PMCID: PMC104969

PMID: 9650953

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https://europepmc.org/articles/pmc104969View

Published (Version of record) Open Access

Abstract

Hepatitis A virus (HAV) immunoassays use cell culture-derived HAV antigen to detect HAV-specific antibodies. The current method of production of HAV antigen in tissue culture is time-consuming and expensive. We previously expressed the HAV open reading frame in recombinant vaccinia viruses (rV-ORF). The recombinant HAV polyprotein was accurately processed and was assembled into subviral particles. These particles were bound by HAV-neutralizing antibodies and were able to elicit antibodies which were detected by commercial immunoassays. The present investigation compared the production of HAV antigen by standard tissue culture methods to the production of HAV antigen with the recombinant vaccinia virus system. In addition, HAV and rV-ORF antigens were assessed for their utility in diagnostic immunoassays. Serum or plasma samples from HAV antibody-positive and antibody-negative individuals were evaluated by immunoassay that used either HAV or rV-ORF antigen. All samples (86 of 86) in which HAV antibody was detected by a commercial enzyme-linked immunosorbent assay (ELISA) also tested positive by the recombinant antigen-based immunoassay (VacRIA). Similarly, all samples (50 of 50) that were HAV antibody negative also tested negative by the VacRIA. The lower limit of detection of HAV antibody was similar among immunoassays with either HAV or rV-ORF antigen. Thus, in the population studied, the sensitivity and specificity of the VacRIA were equivalent to those of the commercial ELISA. Since production of recombinant antigen is faster and less expensive than production of traditional HAV antigen, the development of diagnostic HAV antibody tests with recombinant HAV antigen appears warranted.

Radioimmunoassay

Recombinant Fusion Proteins - biosynthesis

Recombinant Fusion Proteins - immunology

Cell Line

Enzyme-Linked Immunosorbent Assay

Humans

Antibodies, Viral - blood

Vaccinia virus - genetics

Antigens, Viral - immunology

Hepatitis A - immunology

Animals

Hepatitis A Antigens

Sensitivity and Specificity

Antibodies, Viral - immunology

Hepatitis A Virus, Human - growth & development

Antigens, Viral - biosynthesis

Hepatitis A - diagnosis

Hepatitis A Virus, Human - immunology

Details

Title: Subtitle: Recombinant hepatitis A virus antigen: improved production and utility in diagnostic immunoassays
Creators: F D LaBrecque - Department of Internal Medicine, Iowa City Veterans Affairs Medical Center and The University of Iowa, 52242, USA
D R LaBrecque
D Klinzman
S Perlman
J B Cederna
P L Winokur
J Q Han
J T Stapleton
Resource Type: Journal article
Publication Details: Journal of clinical microbiology, Vol.36(7), pp.2014-2018
DOI: 10.1128/JCM.36.7.2014-2018.1998
PMID: 9650953
PMCID: PMC104969
NLM abbreviation: J Clin Microbiol
ISSN: 0095-1137
eISSN: 1098-660X
Publisher: United States
Grant note: RR0059 / NCRR NIH HHS M01 RR000059 / NCRR NIH HHS
Language: English
Date published: 07/1998
Academic Unit: Microbiology and Immunology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Medicine Administration; Infectious Disease (Pediatrics); Internal Medicine
Record Identifier: 9983777352202771

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