1. OnSite
Dengue Rapid Test

2. Overview Dengue: Global impact and diagnostic needs
CTK Biotech: The complete solution to dengue diagnosis
CTK product line: Technical details
Summary and suggested literature

3. WHO: The global burden of disease 2004 update
Dengue is a global health concern endemic to tropical and sub-tropical areas including Africa, Asia and the Americas. 2.5 billion people (40% of the world population) are at risk of infection.
Annual Statistics
million infections reported annually
22 thousand deaths (mainly children)
2012 marked the spread to Europe with the first sustained transmission of dengue fever since the 1920’s
Brazil and Mexico make up 14% percent of cases
WHO: The global burden of disease 2004 update

4. WHO: The global burden of disease 2004 update
Southeast Asia bears a large portion of the global burden with Thailand, Indonesia and India already experiencing fatal outbreaks as peak season approaches.
Estimated 70% of world’s serious dengue cases in were in Asia
2013 figures reported since January are on pace for a record setting Dengue season
WHO: The global burden of disease 2004 update

5. Two of the main reasons for the increase in reported cases, global spread of the virus and rise in the severity of cases reported are:
Environmental factors and urbanization of areas conducive to mosquito reproduction  increased risk of infection because of increased exposure to transmitting mosquitoes
All 4 serotypes have been identified in recent outbreaks; historically only 1 or 2 subtypes were identified in a particular outbreak  increase in secondary infections which are more severe than primary infections

6. Four serotypes of dengue virus exist in the endemic areas of the world
Four serotypes of dengue virus exist in the endemic areas of the world. The virus is arthropod-borne, transmitted by mosquitos.
Human
Aedes aegypti
Gunzman et al. Nature review 2010; Center for Disease Control (CDC)
Dengue is a member of the flavi virus family
Four serotypes exist (DENV 1, 2, 3, 4)
The virus is transmitted by tiger mosquitos (Aedes spp.)
Screening and limiting the number of mosquito bites is critical to managing the spread of the disease.

7. Primary infection results in mild symptoms and confers immunity to subsequent infection by the same serotype. Secondary infections are more severe leading to Dengue hemorrhagic fever.
recovery
1° infection
2° infection
Immunity to 1 serotype
DENV serotype
DENV serotype
Any of the 4 DENV serotypes
DENV serotype different from 1° infection
Disease
Disease
Symptoms:
Mild (Dengue Fever DF)
Similar to other infection (Flu or malaria)
Confers immunity against 1 DENV serotype
Symptoms:
Life-threatening (Dengue hemorrhagic fever DHV
and Dengue shock syndrome DSS)
Antibody enhanced infection
Secondary infection is the most significant individual risk factor for severity of disease.
No vaccine is available & treatment is limited to replenishing fluids and electrolytes.
Therefore, early diagnosis that can distinguish between primary and secondary infections is critical to disease management allowing faster treatment and limit mortality and morbidity rates.

8. Dengue infections can be detected at all stages of disease
Dengue infections can be detected at all stages of disease. Different Ab responses during primary and secondary infections enable differentiation between the two types of infection.
Primary dengue infection
Secondary dengue infection
Disease
onset
Disease
onset
virus
virus
IgG
IgG
IgM
IgM
>90
…>90
days
days
viral antigen
(Day 1-5)
viral antigen
(Day 1-5)
Anti-dengue IgM
(day 3-30)
Anti-dengue IgM low and varying
Anti-dengue IgG
(from day 7-11 on)
Anti-dengue IgG
(from day 3 on)
Adapted from Sa-Ngasanget al. Epidemiol. Infect. (2006), Gunzman et al. Int. Jour. Inf. disease 2004

9. Diagnostic needs to manage dengue
Detection of dengue as early as possible to initiate medical care
Discrimination of dengue from other infections with similar symptoms such as Leptospira, rubella
Detection of all 4 DENV serotypes (DENV 1, 2, 3, 4)
Detection during all phases of a dengue infection
Differentiation between primary and secondary dengue infections

10. Training requirements Application in the field
OnSite Rapid Tests offer many advantages compared to other diagnostic methods: minimal resource requirements, faster time to results, use in unlimited test settings
Virus isolation
RT-PCR
ELISA
OnSite Rapid Test
Sensitivity
high
High-medium
Specificity
Equipment required
medium
low
Training requirements
Application in the field no
yes
Time
days
6-12h
60-75min
15-25min
Direct and indirect methods
1. Direct: Detection of virus (virus isolation, detection of viral RNA by RT-PCR,
detection of viral antigens)
2. Indirect: Serology to detect of anti dengue antibodies
Opportunity
Resources

11. CTK Biotech offers a complete line for the diagnosis of infection with dengue virus at various disease stages.
OnSite Rapid Tests
Dengue IgG/IgM Combo Rapid Test (R0061C)
- detection and differentiation of anti-dengue IgG, IgM
Duo Dengue Ag-IgG/IgM Rapid Test (R0062C)
- detection and differentiation viral antigens and anti-dengue IgG, IgM
Dengue Ag Rapid Test (R0063C)
- detection of DENV antigens at early stage of infection (1-5 days)
Positivia External Controls
Dengue Ag Rapid Test Control Kit (C0063)

12. OnSite Dengue Ag Rapid Test
R0063C

13. OnSite Dengue Ag Rapid Test (R0063C) Test Overview
Intended use
Qualitatively detects dengue Ag
Specimen type
Serum/Plasma/Whole Blood
Reading time
25 minutes
Performance
Relative Sensitivity: 95.6%,
Relative Specificity: 95.5%
Shelf life
18 months
Package size
30 tests

14. OnSite Dengue Ag Rapid Test (R0063C)
Test procedure – as easy as 1, 2, 3
Step 1:
Addition of specimen
Step 2:
Addition of sample diluent
Step 3:
Read Result
Result
1 drop serum/plasma
1 drop sample diluent
25 minutes
2 drops whole blood
1 drop sample diluent

15. OnSite Dengue Ag Rapid Test (R0063C) Interpretation of Results
Negative
Positive
Invalid
A positive Dengue Ag test result indicates early stage, current infection.
Treatment should be started immediately to minimize severity of symptoms.

16. OnSite Dengue Ag Rapid Test
OnSite Dengue Ag Rapid Test (R0063C)
Performance Data
OnSite Dengue Ag Rapid Test
Dengue Ag EIA Test
Positive
Negative
Total 66 3 69 2 43 45 68 46
114
Relative Sensitivity: 95.6%
Relative Specificity: 95.5%
Overall Agreement: 95.6%

17. OnSite Dengue Ag Rapid Test (R0063C)
External evaluations performed by independent reference labs in different geographic regions showed favorable results.
Two leading reference labs in Ecuador and one in Sri Lanka recently performed independent evaluations of the OnSite Dengue Ag Rapid Test.
Key findings in the reports:
100% agreement with the reference test used for evaluation
No cross-reactivity with 7 other infectious disease samples
Highly sensitive and specific

18. OnSite Dengue IgG/IgM Combo Rapid Test
R0061C

19. OnSite Dengue IgG/IgM Combo Rapid Test (R0061C) Intended Use
Qualitatively detects & differentiates IgG & IgM to DENV 1, 2, 3, 4
Antigen used
Proprietary recombinant DENV 1, 2, 3 & 4 Ag
Specimen type
Serum/Plasma/Whole blood
Reading time
25 minutes
IgM Performance
Relative Sensitivity: 96.9%; Relative Specificity: 98.9%
IgG Performance
Relative Sensitivity: 97.3%
Relative Specificity: 99.3%
Shelf life
18 months
Package size
30 tests

20. OnSite Dengue IgG/IgM Combo Rapid Test (R0061C)
Test procedure – as easy as 1, 2, 3
Step 1:
Addition of specimen
Step 2:
Addition of sample diluent
Step 3:
Read Result
Result
5 µL serum/plasma
25 minutes
3 drops sample diluent
5 µL whole blood

21. OnSite Dengue IgG/IgM Combo Rapid Test (R0061C)
Interpretation of Results
IgM Positive
IgG
Positive
IgM, IgG Positive
Negative
Invalid
Explain how each result relates back to the various stages of the disease

22. OnSite Dengue IgG/IgM Combo Rapid Test
OnSite Dengue IgG/IgM Combo Rapid Test (R0061C)
Performance Data for IgM test
OnSite Dengue IgG/IgM Combo Rapid Test
IgM EIA Test
Positive
Negative
Total 31 1 32 3
279
282 34
280
314
Relative Sensitivity: 96.9%
Relative Specificity: 98.9%
Overall Agreement: 98.7%

23. OnSite Dengue IgG/IgM Combo Rapid Test
OnSite Dengue IgG/IgM Combo Rapid Test (R0061C)
Performance Data for IgG test
OnSite Dengue IgG/IgM Combo Rapid Test
IgG EIA Test
Positive
Negative
Total 36 1 37 2
287
289 38
288
326
Relative Sensitivity: 97.3%
Relative Specificity: 99.3%
Overall Agreement: 99.1%

24. OnSite Dengue IgG/IgM Combo Rapid Test (R0061C)
External evaluations performed by independent reference labs in different geographic regions showed favorable results
Reference labs in Ecuador and Taiwan recently performed independent evaluations of the OnSite Dengue IgG/IgM Rapid Test.
Key findings in the reports:
100% agreement with the reference test used for evaluation
Report from Taiwan to be published

25. OnSite Duo Dengue Ag-IgG/IgM Rapid Test (R0062C)
Combines the dengue Ag and IgG/IgM tests in a single cassette
Intended use
Qualitatively detects & differentiates IgG & IgM anti-dengue virus and dengue Ag
Antigen used
Proprietary recombinant DENV 1, 2, 3 & 4 Ag
Specimen type
Serum/Plasma/Whole blood
Reading time
25 minutes
Shelf life
18 months
Package size
30 tests

26. Early Diagnosis for a better life Together, we grow Athenese Dx & CTK

27. Together, we grow Athenese Dx & CTK
Athenese DX contact information:
Module # 407 & 408, 4th Floor,
Ticel Bio Park II, No. 5, CSIR Road,
Taramani, Chennai ,
TamilNadu, India.
Together, we grow Athenese Dx & CTK