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Combination of Epstein-Barr virus scaffold (BdRF1/VCA-p40) and small capsid protein (BFRF3/VCA-p18) into a single molecule for improved serodiagnosis of acute and malignant EBV-driven disease.

Abstract
Current single Epstein-Barr virus (EBV) markers fail to reach 100% sensitivity for serodiagnosis of acute and malignant diseases associated with EBV infection. Previous study had identified immunodominant epitopes of VCA-p40 and VCA-p18, and indicated that these two VCA antigens may have diagnostic value for EBV-related diseases. A recombinant protein of the full-length BdRF1 fused to the immunodominant domain of BFRF3 as 6-his tagged protein in Escherichia coli was developed. The recombinant protein was extracted in 8M urea solution and purified by metal-affinity chromatography yielding a 55 kDa product (VCA-p40+18). VCA-p40+18 blot-strips examined for IgM reactivity in infectious mononucleosis samples yielded 100% sensitivity and specificity, with improved reactivity compared with IgM/VCA-p18-ELISAs. A recent study described a synthetic peptide-based IgA/[EBNA1+VCA-p18]-ELISA (IgA/EBV-ELISA), with a sensitivity of 90% for diagnosing nasopharyngeal carcinoma. Immunoblot analysis of biopsy-confirmed nasopharyngeal carcinoma cases with low or negative IgA/EBV-ELISA showed 100% IgG reactivity to VCA-p40 and VCA-p18 proteins. Evaluation of VCA-p40+18 as an additional marker for screening and diagnosis of nasopharyngeal carcinoma was carried out. The data showed positive IgA/VCA-p40+18 reactivity by ELISA for 63.6% (14 of 22) nasopharyngeal carcinoma samples that were missed by peptide-based IgA/EBV-ELISA, suggested VCA-p40+18 as an improved marker for nasopharyngeal carcinoma serodiagnosis. The VCA-p40+18 may be combined with an EBNA1 synthetic peptide as an antigen mixture in one or separate IgA ELISA for improved nasopharyngeal carcinoma serodiagnosis.
AuthorsJajah Fachiroh, Servi J C Stevens, Sofia M Haryana, Jaap M Middeldorp
JournalJournal of virological methods (J Virol Methods) Vol. 169 Issue 1 Pg. 79-86 (Oct 2010) ISSN: 1879-0984 [Electronic] Netherlands
PMID20621126 (Publication Type: Comparative Study, Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright (c) 2010 Elsevier B.V. All rights reserved.
Chemical References
  • Antibodies, Viral
  • Antigens, Viral
  • Capsid Proteins
  • Epstein-Barr viral capsid antigen
  • Immunoglobulin M
  • Recombinant Fusion Proteins
Topics
  • Antibodies, Viral (blood)
  • Antigens, Viral (genetics, immunology)
  • Biopsy
  • Blotting, Western
  • Capsid Proteins (genetics, immunology)
  • Carcinoma
  • Chromatography, Affinity
  • Epstein-Barr Virus Infections (diagnosis)
  • Escherichia coli (genetics)
  • Herpesvirus 4, Human (genetics, immunology)
  • Humans
  • Immunoglobulin M (blood)
  • Infectious Mononucleosis (diagnosis, virology)
  • Molecular Weight
  • Nasopharyngeal Carcinoma
  • Nasopharyngeal Neoplasms (diagnosis, virology)
  • Recombinant Fusion Proteins (chemistry, genetics, isolation & purification)
  • Sensitivity and Specificity
  • Serologic Tests (methods)
  • Virology (methods)

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