1. Panbio Dengue ELISAs

2. Most appropriate use: Panbio Dengue ELISAs

3. Panbio Dengue ELISAs Dengue IgM Capture ELISA (E-DEN01M)
For the diagnosis of active dengue
Suitable for use in endemic and non-endemic dengue regions
Recommended for use with the Dengue IgG Capture ELISA in regions where secondary dengue is prevalent
Dengue IgG Capture ELISA (E-DEN02G)
For the diagnosis of secondary dengue
For use in dengue endemic regions
Use with the Dengue IgM Capture ELISA to detect and distinguish primary and secondary dengue with a high degree of confidence
Dengue Duo IgM and IgG ELISA (E-DEN01D)
This kit consists of the Dengue IgM Capture ELISA and the Dengue IgG Capture ELISA packaged together for convenience and cost savings.
Ideal for use in regions where primary and secondary dengue are common.
For the differentiation of primary and secondary dengue.

4. Panbio Dengue ELISAs cont.
Dengue IgG Indirect ELISA (E-DEN01G)
For detecting past or active dengue
Suitable for seroepidemiological studies

5. Positioning Laboratory testing
Recommend test based on location and patient types
i.e. endemic region v non-endemic region
Is differentiation between primary and secondary important?
Diagnosis of infection v seroepidemiological screening
Emphasize Panbio: No 1 in dengue fever detection
Numerous publications to support this

6. Panbio Dengue IgG Indirect ELISA
Specific serum antibodies combine with dengue 1-4
antigens attached to the polystyrene
surface of the microwells
Washing removes residual serum
Peroxidase-conjugated anti-human specific
immunoglobulin is added
The colourless substrate,
tetramethylbenzidine/hydrogen peroxide (TMB /
H2O2) is hydrolysed to a blue chromogen
Stopping the hydrolytic reaction
with acid turns the TMB yellow
Colour development indicates the presence
dengue antibodies in the test sample

7. Dengue IgG Indirect ELISA
Anti-human IgG HRP conjugated
Patient’s dengue specific IgG
Dengue antigen coated plate

8. Panbio Dengue IgM & IgG Capture ELISAs

9. Dengue IgM Capture ELISA
Dengue antigen-Mab complex
IgM in patient’s serum
Anti-human IgM coated plate
Dengue IgG Capture ELISA is essentially the same except anti-human IgG antibodies are coated on the plate and therefore IgG in the patient’s serum is captured.

10. Interpretation: Capture ELISAs
IgM positive / IgG negative
Serology indicates the probability that the patient has current or recent primary dengue infection.
IgM positive / IgG positive
Serology indicates the probability that the patient has current or recent secondary dengue infection.
IgM negative (equivocal) / IgG positive
In secondary dengue IgM may be negative in up to 75% of patients, depending on the stage of disease when the sample was collected.

11. Interpretation cont: Capture ELISAs
IgM equivocal / IgG negative
Serology indicates that the patient may have current or recent primary dengue infection.
If clinical signs and symptoms persist recommend that a second serum sample be collected and tested.
IgM positive / IgG equivocal
Serology indicates the probability that the patient has current or recent primary dengue infection.
Equivocal IgG suggests that the patient may have current or recent secondary dengue infection.
Patient should be monitored.

12. Interpretation cont: Capture ELISAs
IgM negative / IgG negative
Serology indicates the probability that the patient does not have current or recent dengue infection.
Note: Serum samples collected early in infection may yield a negative serological test result.
If a differential diagnosis can not be established and clinical signs and symptoms persist, recommend that a second serum be collected and tested.
IgM negative / IgG equivocal
Equivocal IgG suggests that the patient may have current or recent secondary dengue infection.
Patient should be monitored.
If clinical signs and symptoms persist recommend that a second serum sample be collected and tested.

13. Interpretation cont: Capture ELISAs
IgM positive / IgG positive
Suggestive of an active dengue infection. Could be either primary or secondary. Use the Dengue IgG Capture ELISA if distinction between primary and secondary dengue is necessary.
IgM negative / IgG positive
Suggestive of a past infection. Secondary dengue infection cannot be ruled out as IgM levels in secondary dengue may be undetectable. If the patients IgG result is >40 Panbio Units, it is likely that the patient has a secondary infection.

14. Performance: Capture ELISAs
Serological sensitivity and specificitya
High sensitivity
Some patients exhibit no IgM in a secondary infection. Therefore detection of high IgG levels to improve the sensitivity is recommended
High specificity

15. Performance: IgM Capture ELISA
Cross reactivity study
94.8% specificity

16. Performance: IgG Capture ELISA
Cross reactivity study
96.8% specificity

17. Publications: Panbio Dengue ELISAs
Vaughn et al (1999). Rapid serological diagnosis of dengue virus infection using a commercial capture ELISA that distinguishes primary and secondary dengue infections. Am.J.Trop.Med.Hyg., 60:
Lam & Devine (1998). Evaluation of capture ELISA and rapid immunochromatographic test for the determination of IgM and IgG antibodies produced during dengue infection. Clin.Diagn.Virol. 10:75-81.
Chew et al. (1998). Evaluation of commercial capture ELISA for detection of immunoglobulin M and G antibodies produced during dengue infection. Clin.Diagn.Lab.Immunol. 5:7-10.
Cuzzubbo et al. (1997). Commercial assays for the serological diagnosis of dengue infection. In Arbovirus Research in Australia 7:56-60.
Phillips et al. (1997). Flavivirus diagnostic assays: the Queensland perspective. In Arbovirus Research in Australia 7:
Cuzzubbo et al. (1998). Detection of specific antibodies in saliva during dengue infection. J. Clin. Microbiol. 36:
Allwin et al. (1998). Abstract. Assessment of flavivirus-specific antibody cross-reactivity after vaccination against TBE and yellow fever. Eur. Conf. Clin. Vir., Hamburg.
Available upon request

18. Commonly Asked Questions about Panbio dengue tests...
Q In the Panbio Dengue Duo ELISA and Dengue Rapid Tests, why don’t symptomatic donors from endemic areas show elevation of IgG?
A IgG Cutoff set above the level in Primary and Past Dengue.

19. Commonly Asked Questions about Panbio dengue tests...
Q How do the cutoffs of these tests compare with haemagglutination-inhibition assays?
A IgG Cutoff = HAI Titre for 2° Dengue (e.g. HAI 1:2560).

20. Commonly Asked Questions about Panbio dengue tests...
Q Do I need to use both IgM and IgG for the diagnosis of dengue infection? Why can’t I just use IgM?
A Some cases of secondary dengue do not show detectable IgM.

21. References Dengue IgM Capture ELISA Instructions for Use. Rev 08/12/04
Dengue IgG Capture ELISA Instructions for Use. Rev 20/10/03

22. Panbio Dengue: The complete range
ELISA kits
Dengue IgG Indirect E-DEN01G
Dengue IgM Capture E-DEN01M
Dengue IgG Capture E-DEN02G
Dengue Duo Capture IgG & IgM E-DEN01D
Rapid kits
Dengue Rapid Strip test R-DEN02D
Dengue Rapid Cassette R-DEN03D