OnSite Dengue Rapid Test

Overview• Dengue: Global impact and diagnostic needs

• CTK Biotech: The complete solution to dengue diagnosis

• CTK product line: Technical details

•Summary and suggested literature

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.

WHO: The global burden of disease 2004 update

Annual Statistics• 50-100 million infections reported annually

• 22 thousand deaths (mainly children)

Southeast Asia bears a large portion of the global burden with Thailand, Indonesia and India already experiencing fatal outbreaks as peak season approaches.

WHO: The global burden of disease 2004 update

• Estimated 70% of world’s serious dengue cases in 2012 were in Asia

• 2013 figures reported since January are on pace for a record setting Dengue season

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:

1)Environmental factors and urbanization of areas conducive to mosquito reproduction increased risk of infection because of increased exposure to transmitting mosquitoes

2)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

Four serotypes of dengue virus exist in the endemic areas of the world. The virus is arthropod-borne, transmitted by mosquitos.

HumanAedes aegypti

• 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.)

Gunzman et al. Nature review 2010; Center for Disease Control (CDC)

Screening and limiting the number of mosquito bites is critical to managing the spread of the disease.

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.

• 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

• Any of the 4 DENV serotypes • DENV serotype different from 1° infectionDENV serotypeDENV serotype

Disease Disease

1° infection 2° infectionrecovery

Immunity to 1 serotype

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.

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.

Disease onset

viral antigen(Day 1-5)

IgGIgM

0 1 2 3 4 5 6 7 8 9 10 30 .. 60...>90days

IgG

Primary dengue infection Secondary dengue infection

IgM

virusvirus

Disease onset

days

Anti-dengue IgG(from day 7-11 on)

viral antigen(Day 1-5)

Anti-dengue IgM(day 3-30)

Adapted from Sa-Ngasanget al. Epidemiol. Infect. (2006), Gunzman et al. Int. Jour. Inf. disease 2004

0 1 2 3 4 5 6 7 8 9 10 30 .. 60…>90

Anti-dengue IgM low and varying

Anti-dengue IgG(from day 3 on)

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

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 ELISAOnSite Rapid

Test

Sensitivity high highHigh-

mediumHigh-

medium

Specificity high highHigh-

mediumHigh-

medium

Equipment required

high high medium low

Training requirements

high high medium low

Application in the field

no no no yes

Time days 6-12h 60-75min 15-25min

OpportunityResources

CTK Biotech offers a complete line for the diagnosis of infection with dengue virus at various disease stages.

OnSite Rapid TestsDengue 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 ControlsDengue Ag Rapid Test Control Kit (C0063)

OnSite Dengue Ag Rapid TestR0063C

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

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

1 drop serum/plasma

2 drops whole blood

1 drop sample diluent

1 drop sample diluent

25 minutesResult

Step 3: Read Result

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.

OnSite Dengue Ag Rapid Test (R0063C) Performance Data

OnSite Dengue Ag Rapid Test

Dengue Ag EIA Test

Positive Negative Total

Positive 66 3 69

Negative 2 43 45

Total 68 46 114Relative Sensitivity: 95.6%

Relative Specificity: 95.5%

Overall Agreement: 95.6%

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

OnSite Dengue IgG/IgM Combo Rapid TestR0061C

OnSite Dengue IgG/IgM Combo Rapid Test (R0061C) Intended Use

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

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

5 µL serum/plasma

5 µL whole blood

3 drops sample diluent

25 minutesResult

Step 3: Read Result

OnSite Dengue IgG/IgM Combo Rapid Test (R0061C) Interpretation of Results

Negative IgM Positive

InvalidIgM, IgG Positive

IgG Positive

OnSite Dengue IgG/IgM Combo Rapid Test (R0061C) Performance Data for IgM test

Relative Sensitivity: 96.9%

Relative Specificity: 98.9%

Overall Agreement: 98.7%

OnSite Dengue IgG/IgM Combo Rapid Test

IgM EIA Test Positive Negative Total

Positive 31 1 32

Negative 3 279 282

Total 34 280 314

OnSite Dengue IgG/IgM Combo Rapid Test (R0061C) Performance Data for IgG test

Relative Sensitivity: 97.3%

Relative Specificity: 99.3%

Overall Agreement: 99.1%

OnSite Dengue IgG/IgM Combo Rapid Test

IgG EIA Test Positive Negative Total

Positive 36 1 37

Negative 2 287 289

Total 38 288 326

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

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

Together, we grow Athenese Dx & CTK

Early Diagnosis for a better life

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 - 600113,TamilNadu, India.