Transcript Document

EBV Competitors
Meridian Bioscience
Premier EBV EA
Premier EBV VCA IgG
Premier EBV VCA IgM
Premier EBV EBNA-1 IgG
Biotest Diagnostics
Biotest Anti-EBV Recombinant EA-IgM ELISA
Biotest Anti-EBV Recombinant EA-IgG ELISA
Biotest Anti-EBV Recombinant EBNA-IgG ELISA
CE Marked
CE Marked
CE Marked
CE Marked
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
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
• 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)
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
Panbio vs Trinity Biotech
Sensitivity
Specificity
Trinity EBV VCA IgG*
91.5
51.2
Panbio EBV VCA IgG
95.5
87.2
Trinity EBV VCA IgM*
87
88.3
Panbio EBV VCA IgM
100
92.3
* 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.
Panbio vs Meridian
Sensitivity
Specificity
Meridian EBV VCA IgM*
94.3
94.8
Panbio EBV VCA IgM
100
92.3
* 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.
www.panbio.com
Epstein-Barr Virus
Competitor Analysis
© 2004 PANBIO Limited. All Rights
Reserved Rev 2004/05
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,
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
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%)
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.
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
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)
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
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
DiaSorin
ETI-VCA-G, ETI-EBV-M-reverse, ETI-EBNA-G, ETIEA-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
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
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%)
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 cterminal 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%)
Comparison studies
1.
2.
3.
4.
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.
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.
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.
2. PANBIO Inhouse data
(2002) cont.
VCA IgG Kits
Specificity
PANBIO (Stellar) IFA
100%
PANBIO ELISA (gp125)
95%
Diasorin ELISA
79%
Trinity ELISA
100%
Zeus ELISA
97%
DiaMedix ELISA
100%
2. PANBIO Inhouse data
(2002) cont.
VCA IgG Kits
Acute
samples
Convalescen
t samples
Overall
PANBIO (Stellar)
IFA
100%
100%
100%
PANBIO ELISA
91%
96%
94%
Diasorin ELISA
71%
94%
87%
Trinity ELISA
26%
88%
69%
Zeus ELISA
44%
96%
79%
DiaMedix ELISA
42%
94%
79%
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.
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%.
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.