Panbio Dengue ELISAs

Panbio Dengue ELISAs

Most appropriate use: Panbio Dengue ELISAs

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.

Panbio Dengue ELISAs cont.• Dengue IgG Indirect ELISA (E-DEN01G)

– For detecting past or active dengue

– Suitable for seroepidemiological studies

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

Specific serum antibodies combine with dengue 1-4 antigens attached to the polystyrenesurface 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 reactionwith acid turns the TMB yellow

Colour development indicates the presencedengue antibodies in the test sample

Panbio Dengue IgG Indirect ELISA

Dengue IgG Indirect ELISA

Dengue antigen coated plate

Patient’s dengue specific IgG

Anti-human IgG HRP conjugated

Panbio Dengue IgM & IgG Capture ELISAs

Dengue IgM Capture ELISA

Anti-human IgM coated plate

IgM in patient’s serum

Dengue antigen-Mab complex

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.

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• Serology indicates the probability that the patient has current

or recent secondary dengue infection.– In secondary dengue IgM may be negative in up to 75% of patients,

depending on the stage of disease when the sample was collected.

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. – If clinical signs and symptoms persist recommend that a second

serum sample be collected and tested.

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.

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.

Performance: Capture ELISAsSerological sensitivity and specificitya

Some patients exhibit no IgM in a secondary infection. Therefore detection of high IgG levels to improve the sensitivity is recommended

High sensitivityHigh specificity

Performance: IgM Capture ELISA Cross reactivity study

94.8% specificity

Performance: IgG Capture ELISA

96.8% specificity

Cross reactivity study

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., 60:693-698.

• 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 7:208-211.

• Cuzzubbo et al. (1998). Detection of specific antibodies in saliva during dengue infection. J. Clin. Microbiol. 36:3737-3739.

• 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

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.

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).

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.

References1. Dengue IgM Capture ELISA Instructions for Use. Rev

08/12/04

2. Dengue IgG Capture ELISA Instructions for Use. Rev 20/10/03

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

Panbio Dengue: The complete range

www.panbio.com