Serodiagnosis of Epstein-Barr Virus Infection by Using Recombinant Viral Capsid Antigen Fragments and Autologous Gene Fusion

Using recombinant 15- to 30-kDa fragments and fusion with glutathione S-transferase (GST), we investigated the seroreactivity of three large structural proteins of Epstein-Barr virus (EBV), p150 (BcLF1, capsid), p143 (BNRF1, tegument), and gp125 (BALF4, membrane) in Western blots. None of 13 fragmen...

Fuld beskrivelse

Na minha lista:
Bibliografiske detaljer
Main Authors: Hinderer, Walter, Lang, Dieter, Rothe, Markus, Vornhagen, Rolf, Sonneborn, Hans H., Wolf, Hans
Format: Artigo
Sprog:en
Udgivet: American Society for Microbiology 1999
Fag:
Virology
Online adgang:https://ncbi.nlm.nih.gov/pmc/articles/PMC85538/
https://ncbi.nlm.nih.gov/pubmed/10488185
Tags: Tilføj Tag
Ingen Tags, Vær først til at tagge denne postø!
id pubmed-85538
record_format dspace
spelling pubmed-855382002-05-02 Serodiagnosis of Epstein-Barr Virus Infection by Using Recombinant Viral Capsid Antigen Fragments and Autologous Gene Fusion Hinderer, Walter Lang, Dieter Rothe, Markus Vornhagen, Rolf Sonneborn, Hans H. Wolf, Hans J Clin Microbiol Virology Using recombinant 15- to 30-kDa fragments and fusion with glutathione S-transferase (GST), we investigated the seroreactivity of three large structural proteins of Epstein-Barr virus (EBV), p150 (BcLF1, capsid), p143 (BNRF1, tegument), and gp125 (BALF4, membrane) in Western blots. None of 13 fragments tested, however, was qualified for diagnostic application. In contrast, the two small viral capsid antigens (VCA), p18 (BFRF3) and p23 (BLRF2), demonstrated sensitive (100%) EBV-specific immunoglobulin G (IgG) reactivities. While p18 additionally showed maximum sensitivity for IgM detection, the IgM sensitivity of p23 was restricted (44%). An autologous fusion protein, p23-p18, which consists N-terminally of full-length p23, followed by the carboxy half of p18, was constructed. This antigen was subjected to indirect VCA enzyme-linked immunosorbent assays (ELISAs), for IgG and IgM, as well as to a μ-capture (μc) IgM ELISA. All assays were found to be 100% specific when EBV-negative sera were tested. Using sera from previously infected individuals, the p23-p18 fusion revealed an improved IgG sensitivity of 99% compared to sensitivities of 97 and 93% for the single antigens p18 and p23, respectively. The sensitivity and specificity of the indirect IgM ELISA with samples of primary and past infections, respectively, were 100%. The μc principle for IgM overcame completely the interference by rheumatoid factors. Compared to the specificity of the indirect IgM version, the specificity with sera collected from rheumatoid arthritis patients increased from 48 to 100%. In summary, the p23-p18 IgG and μc IgM ELISAs showed excellent performances and are promising new diagnostic tests for the detection of EBV-specific antiviral capsid antibodies. American Society for Microbiology 1999-10 /pmc/articles/PMC85538/ /pubmed/10488185 Text en Copyright © 1999, American Society for Microbiology
institution US National Library of Medicine
collection PubMed Central
language en
format Article
topic Virology
spellingShingle Virology
Hinderer, Walter
Lang, Dieter
Rothe, Markus
Vornhagen, Rolf
Sonneborn, Hans H.
Wolf, Hans
Serodiagnosis of Epstein-Barr Virus Infection by Using Recombinant Viral Capsid Antigen Fragments and Autologous Gene Fusion
description Using recombinant 15- to 30-kDa fragments and fusion with glutathione S-transferase (GST), we investigated the seroreactivity of three large structural proteins of Epstein-Barr virus (EBV), p150 (BcLF1, capsid), p143 (BNRF1, tegument), and gp125 (BALF4, membrane) in Western blots. None of 13 fragments tested, however, was qualified for diagnostic application. In contrast, the two small viral capsid antigens (VCA), p18 (BFRF3) and p23 (BLRF2), demonstrated sensitive (100%) EBV-specific immunoglobulin G (IgG) reactivities. While p18 additionally showed maximum sensitivity for IgM detection, the IgM sensitivity of p23 was restricted (44%). An autologous fusion protein, p23-p18, which consists N-terminally of full-length p23, followed by the carboxy half of p18, was constructed. This antigen was subjected to indirect VCA enzyme-linked immunosorbent assays (ELISAs), for IgG and IgM, as well as to a μ-capture (μc) IgM ELISA. All assays were found to be 100% specific when EBV-negative sera were tested. Using sera from previously infected individuals, the p23-p18 fusion revealed an improved IgG sensitivity of 99% compared to sensitivities of 97 and 93% for the single antigens p18 and p23, respectively. The sensitivity and specificity of the indirect IgM ELISA with samples of primary and past infections, respectively, were 100%. The μc principle for IgM overcame completely the interference by rheumatoid factors. Compared to the specificity of the indirect IgM version, the specificity with sera collected from rheumatoid arthritis patients increased from 48 to 100%. In summary, the p23-p18 IgG and μc IgM ELISAs showed excellent performances and are promising new diagnostic tests for the detection of EBV-specific antiviral capsid antibodies.
author Hinderer, Walter
Lang, Dieter
Rothe, Markus
Vornhagen, Rolf
Sonneborn, Hans H.
Wolf, Hans
author_facet Hinderer, Walter
Lang, Dieter
Rothe, Markus
Vornhagen, Rolf
Sonneborn, Hans H.
Wolf, Hans
author_sort Hinderer, Walter
title Serodiagnosis of Epstein-Barr Virus Infection by Using Recombinant Viral Capsid Antigen Fragments and Autologous Gene Fusion
title_short Serodiagnosis of Epstein-Barr Virus Infection by Using Recombinant Viral Capsid Antigen Fragments and Autologous Gene Fusion
title_full Serodiagnosis of Epstein-Barr Virus Infection by Using Recombinant Viral Capsid Antigen Fragments and Autologous Gene Fusion
title_fullStr Serodiagnosis of Epstein-Barr Virus Infection by Using Recombinant Viral Capsid Antigen Fragments and Autologous Gene Fusion
title_full_unstemmed Serodiagnosis of Epstein-Barr Virus Infection by Using Recombinant Viral Capsid Antigen Fragments and Autologous Gene Fusion
title_sort serodiagnosis of epstein-barr virus infection by using recombinant viral capsid antigen fragments and autologous gene fusion
publisher American Society for Microbiology
publisher_facet American Society for Microbiology
publishDate 1999
url https://ncbi.nlm.nih.gov/pmc/articles/PMC85538/
https://ncbi.nlm.nih.gov/pubmed/10488185
_version_ 1759028708147986432

Lignende værker