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EBV Competitors

EBV Competitors. Meridian Bioscience Premier EBV EA CE Marked Premier EBV VCA IgG CE Marked Premier EBV VCA IgM CE Marked Premier EBV EBNA-1 IgG CE Marked Biotest Diagnostics Biotest Anti-EBV Recombinant EA-IgM E LISA Biotest Anti-EBV Recombinant EA-IgG E LISA

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EBV Competitors

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  1. EBV Competitors Meridian Bioscience Premier EBV EA CE Marked Premier EBV VCA IgG CE Marked Premier EBV VCA IgM CE Marked Premier EBV EBNA-1 IgG CE Marked Biotest Diagnostics Biotest Anti-EBV Recombinant EA-IgM ELISA Biotest Anti-EBV Recombinant EA-IgG ELISA Biotest Anti-EBV Recombinant EBNA-IgG ELISA

  2. EBV Competitors Dade Behring – Enzygnost Enzygnost® Anti-EBV/IgG Enzygnost® Anti-EBV/IgG Enzygnost® Anti-EBV/IgG Avidity Enzygnost® Anti-EBV/IgM Enzygnost® Anti-EBV/IgM Trinity Biotech CAPTIATM Epstein Barr Virus (EBNA-1) IgG Epstein Barr Virus (EBNA-1) IgM Epstein Barr Virus (EA-D) IgG Epstein Barr Virus (VCA) P-18 IgG Epstein Barr Virus VCA IgM

  3. EBV Competitors Vironostika (Biomerieux) Enzygnost® Anti-EBV/IgG Enzygnost® Anti-EBV/IgG Enzygnost® Anti-EBV/IgG Avidity Enzygnost® Anti-EBV/IgM Enzygnost® Anti-EBV/IgM Trinity Biotech CAPTIATM Epstein Barr Virus (EBNA-1) IgG Epstein Barr Virus (EBNA-1) IgM Epstein Barr Virus (EA-D) IgG Epstein Barr Virus (VCA) P-18 IgG Epstein Barr Virus VCA IgM

  4. VironostikaVCA IgM/IgG/EBNA EIA (Organon Teknika) • Clin-ELISA EBV (DiaSorin) • Zeus (EBV-VCA IgG, EBV-VCA IgM) • Diamedix (EBV-VCA IgG, EBV-VCA IgM, EBNA-1 IgG, EBNA-1 IgM, EBV-EA-D IgG, EBV-EA-D IgM)

  5. Panbio vs Trinity Biotech • Trinity Biotech • Indirect • Incubation times of 20 minutes • Different methods for VCA IgM and IgG kits • 7 wells required for control samples • Panbio • Indirect • Incubation times of 30 minutes • All Panbio EBV kits have the same procedure • Panbio kits only require 5 wells for control samples

  6. Panbio vs Trinity Biotech * Dickeson, D.J. et al. (2001). Evaluation of commercial E.I.A. kits for the detection of Epstein Barr virus viral capsid antigen IgG and IgM. Poster P4.9 Australian Society for Microbiology, Annual Scientific Meeting, Perth, Australia, October 2001.

  7. Panbio vs Meridian * Dickeson, D.J. et al. (2001). Evaluation of commercial E.I.A. kits for the detection of Epstein Barr virus viral capsid antigen IgG and IgM. Poster P4.9 Australian Society for Microbiology, Annual Scientific Meeting, Perth, Australia, October 2001.

  8. www.panbio.com

  9. Epstein-Barr VirusCompetitor Analysis © 2004 PANBIO Limited. All Rights Reserved Rev 2004/05

  10. Competitor ELISAs • Captia Select VCA IgM/IgG, EBNA IgG/IgM, EA-D IgG EIA (Trinity/Clark) • Enzygnost EBV IgM/IgG EIA (Dade Behring) • Vironostika VCA IgM/IgG/EBNA EIA (Organon Teknika) • Biotest EBV IgM/EA IgG/EBNA IgG EIA • Meridian (Gull) VCA IgM/IgG/EBNA EIA • Clin-ELISA EBV (DiaSorin) • Zeus (EBV-VCA IgG, EBV-VCA IgM) • Diamedix (EBV-VCA IgG, EBV-VCA IgM, EBNA-1 IgG, EBNA-1 IgM, EBV-EA-D IgG, EBV-EA-D IgM)

  11. CAPTIA Select(Trinity/Clark) • VCA IgM/IgG, EBNA IgG/IgM & EA-D IgG ELISAs • VCA IgM: capture • VCA IgG, EBNA, EA-D: indirect • Antigens • VCA IgG Ag: 47kDa fusion pr of 53 amino acids from the c-terminal half of p18 • VCA IgM Ag: gp 125 • EA-D: rec EA-D • EBNA: rec EBNA-1

  12. CAPTIA Select cont. • Disadvantages • Panel lacks specificity, especially with sera from patients with no previous EBV infection (Bruu et al) • 7 wells required for control samples • Different methods for VCA IgG and IgM kits • Short incubation times of 20’ – difficult for some automated systems • Panbio in-house studies have shown the VCA IgG kit lacks sensitivity (69%)

  13. Enzygnost (Dade Behring) • EBV IgM/IgG ELISAs • Indirect; Qualitative and quantitative • Antigen: combo of EBV-specific epitopes; control Ag • Sensitivity 99%; Specificity 100% (Bruu et al., 2000) • Disadvantages: • Unable to distinguish between VCA IgG & EBNA Abs. • The IgM ELISA gives many equivocal results. • Time consuming due to complicated dilution steps.

  14. Vironostika(Organon Teknika) VCA IgM/IgG & EBNA IgG ELISAs • VCA IgM: capture • Mab anti-VCA p18 labeled with peroxidase • Synthetic VCA p18 peptide in conj. diluent • VCA IgG & EBNA IgG: indirect • VCA Ag: synthetic p18 peptide • Qualitative and semiquantitative detection • Sensitivity 95%; Specificity 89% (Bruu et al., 2000) • Disadvantages • lacks specificity

  15. Biotest Diagnostics EA-D IgM/IgG & EBNA IgG ELISAs • Indirect; Qualitative & semiquantitative • EA-D IgG/IgM • Rec. Antigens: EA-D-p54 & EA-p138 • Total inc. time 1h45min for IgM; 1h15min for IgG • EBNA IgG • Rec. Antigen: EBNA-1 p-172 • Total inc. time 1hr15mins • Disadvantages: • Low specificity of EA-D IgM (64%)(Svahn et al,1997) • Low specificity of panel (Bruu et al., 2000)

  16. Meridian/Gull VCA IgM/IgG & EBNA IgG ELISAs • Indirect; Qualitative • Antigens • VCA Ag: purified EBV VCA gp125 • EBNA Ag: recombinant EBNA-1 minus Gly-Ala repeat • Sensitivity 96%; Specificity 90% (Bruu et al., 2000) • Total inc. time 1h30min • Colour change sample diluent • Disadvantage • Preparation of substrate

  17. DIESSE VCA IgM/IgG; EA-D IgM/IgG; EBNA IgG • Indirect; Qualitative • Antigens • VCA Ag: purified EBV Ag • EBNA Ag: rec EBNA-1 • EA-D IgM/IgG: recombinant polypeptides produced in E. coli containing epitopes of the proteins p138 and p54. • Total inc. time 1h45mins • Disadvantages • Instability of reagents after opening (plate stable, wash buffer, serum diluent) • Sample diluent must be diluted prior to use

  18. DiaSorin ETI-VCA-G, ETI-EBV-M-reverse, ETI-EBNA-G, ETI-EA-G • VCA, EBNA & EA IgG: Indirect, qualitative or quantitative, 2h30mins • VCA IgM: Capture, qualitative or quantitative, 2h30mins • VCA Ag: purified EBV Ag • EBNA Ag: rec EBNA-1 • EA-D IgM/IgG: recombinant polypeptides produced in E. coli containing epitopes of the proteins p138 and p54. • Antigens • VCA IgM & IgG: VCA p18 • EBNA IgG: EBNA-1 synthetic peptide • EA: EA-D rec 47kDa polypeptide

  19. DiaSorin cont. • Disadvantages • Conj must be diluted prior to use & can’t be stored • Opened plate can only be stored for 8 weeks • Control sera are supplied ready to use – not treated in the same manner as patients • 9 wells are required for blank and controls when run quantitatively, 6 wells required for blank and controls when run qualitatively

  20. Zeus EBV-VCA IgG, EBV-VCA IgM • Indirect, 50 min total inc time • Antigen • VCA IgG: VCA Ag from P3H3 induced cells • VCA IgM: affinity purified 125kDa capsid peptide • Disadvantages • Short incubation times of 20’-difficult for some automated systems • 6 wells required for controls and blank • PANBIO in-house studies have shown the VCA IgG kit lacks sensitivity (77%)

  21. Diamedix EBV-VCA IgG, EBV-VCA IgM, EBNA-1 IgG, EBNA-1 IgM, EBV-EA-D IgG, EBV-EA-D IgM • Indirect, qualitative, 90min • Antigens • VCA IgG & IgM: 47kDa fusion pr of 53 amino acids from the c-terminal half of p18 • EBNA: rec EBNA-1 • EA: purified rec EA-D • Disadvantages • Serum dilution different for VCA IgM and IgG kits • PANBIO in-house studies have shown the VCA IgG kit lacks sensitivity (79%)

  22. Comparison studies • Dickeson et al. ICPMR, Westmead Hospital. (2001) Evaluation of commercial EIA kits for the detection of Epstein-Barr virus viral capsid antigen IgG and IgM. Poster P4.9. ASM Annual Scientific Meeting, Perth, WA 4 Octover 2001. • PANBIO Inhouse data (2002) Comparison of commercially available Epstein-Barr virus ELISA and IFA kits. BREBV6 Rev2002/10. • Cannone et al. (2002) A comparison of three commercially available Epstein-Barr virus VCA-p18 IgG ELISAs. Poster presented at ASM Annual Scientific Meeting, Melbourne, VIC, October 2002. • Gibb and McGoldrick. (2000) Serological Assays for EBV: Comparison of the Gulll and PANBIO EBV VCA IgG, EBV VCA IgM and EBNA IgG Assays. PANBIO News, Issue 18.

  23. 1. Dickeson et al. (2001)Conclusions • Compared VCA IgG & IgM ELISA kits from Biotec, Diesse, Gen-Bio, Gull/Meridian, PANBIO (gp125), Zeus, Trinity Biotech & IBL. • The best EIA kits for VCA IgG were Biotec & PANBIO. • The best kits for VCA IgM were GenBio, PANBIO and Zeus. • GenBio had complicated result calculations while Zeus had short incubation times. • PANBIO kit with PPV and NPV above 95% were most suitable for routine testing.

  24. 2. PANBIO Inhouse data (2002) • Serum samples characterised using Gull/Meridian IFA • PANBIO (Stellar) VCA IgG IFA kits demonstrated 100% agreement with the Meridian results • The PANBIO VCA (gp125) IgG ELISA performed the best out of the kits tested. F-value (sum of the sensitivity and specificity) was the highest.

  25. 2. PANBIO Inhouse data (2002) cont.

  26. 2. PANBIO Inhouse data (2002) cont.

  27. 3. Cannone et al. (2002) • The PANBIO VCA-p18 IgG ELISA was compared to the Trinity Biotech and Diasorin VCA IgG ELISAs. • The PANBIO kit performed the best in terms of combined specificity and sensitivity for past infection. • The Trinity kit demonstrated a sensitivity of only 69.9% for past infections.

  28. 4. Gibb and McGoldrick. (2000) • The PANBIO VCA (gp125) IgG and IgM and EBNA IgG ELISA kits were compared to the equivalent Gull ELISAs. • The Gull and PANBIO products performed similarly with all kits achieving sensitivities and specificities >96%.

  29. References 1. van Grunsven, W.M.J., W.J.M. Spaan and J.M. Middeldorp. (1994) Localization and diagnostic application of immunodominant domains of the BFRF3-encoded Epstein-Barr virus capsid protein. 2. van Grunsven, W.M.J., A. Nabbe and J.M. Middeldorp. (1993) Identification and molecular characterization of two diagnostically relevant market proteins of the Epstein-Barr virus capsid antigen. J. Med. Virol. 40:161-169. 3. Hinderer, W., D. Lang, M. Rothe, R. Vornhagen, H.H. Sonneborn and H. Wolf. (1999) Serodiagnosis of Epstein-Barr virus infection by using recombinant viral capsid antigen fragments and autologous gene fusion. J. Clin. Microbiol. 37:3239-3244. 4. Bauer, G. (2001) Simplicity through complexity: immunoblot with recombinant antigens as the new gold standard in Epstein-Barr virus serology. Clin. Path. 47:223-230.

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