QUALITATIVE DETECTION OF DENGUE NS1 ANTIGEN & IgG/IgM ANTIBODY

Lateral flow immunochromatographic assay

Principle of the test Dengue NS1 and IgG/IgM antibody

Enveloped, Single-stranded, Positive-sense RNA virus, arthropod-borne

Dengue virus characteristic

50 nanometers

Diameter

Flaviviridae

FAMILY

Stegomyia

SUBFAMILY

Flavivirus

GENUS

Envelope protein

For attachment and fusion

Colloidal gold nanoparticles

-Contains antibody

-Causes color

-Embedded in the conjugate pad

Non Structural protein

Viral replications

Non structural 1 antigen

Antigen being detected

Active replication

NS1 positive

Rough endoplasmic reticulum

Organelle of the life cycle

Envelope protein

DENV virion

DC SIGN, heparan sulfate, FcγR

Host cell receptor

Mononuclear phagocytic system

Primary target cells

Receptor-mediated endocytosis

Process of entry

Aedes aegypti

Most significant epidemic vector in tropical and subtropical

Aedes aegypti

-Primary carrier of the dengue

-Yellow fever mosquito

-African mosquito

-Color brown

Aedes aegypti shape of the thorax lines

Four lines of white scales in the shape of a lyre on the dorsal side

Perpendicular white scale in every single segment

Aedes aegypti scales of the

Aedes albopictus

Shape of the thorax lines: One single white scale

Has one similar scale but thinner, with triangular spots

Aedes aegypti scales of the Abdomen

Liver

Major site of DENV replication

Febrile, Critical, Recovery

Clinical phases of dengue

Without warning signs, With warning signs, Severe dengue

Severity of dengue

Severe abdominal pain, Persistent vomiting, Mucosal bleeding, Lethargy or restlessness

Warning signs

Dengue fever

-High fever and flu-like symptoms including joint muscle pain, rash and headaches

-Typically self limited

Dengue hemorrhagic fever

-Increased plasma leakage(resulting in hemoconcentration) severe bleeding thrombocytopenia(low platelet count)

-With symptoms like: Severe fever hepatomegaly hemorrhagic manifestations.

Dengue shock syndrome

-Most severe form

-Characterized by circulatory failure(shock) from severe plasma leakage.

Dengue Hemorrhagic Fever and Dengue Shock Syndrome

Severe for and could result in liver involvement

Anti-Dengue virus IgM

-Starts to appear 3 days after initial exposure.

-Remains in circulation for 30-60 days.

7 days

Anti-Dengue virus IgG rises around at

2-3 weeks

Anti-Dengue virus IgG peaks at

Antibody Dependent Enhancement

-Mainly mediated by IgG

-Facilitates viral uptake

-Enhances viral replications

NS1

Antigen

IgG and IgM

Antibody

Anti-dengue IgG/IgM rapid test

Left side of the kit

Non structural 1 Dengue Test

Right side of the kit

100 microliters

Specimen ul of NS1 Dengue Test

10 microliters

Specimen ul of Anti-dengue IgM/IgG Test

90 microliters

Buffer ul of Anti-dengue IgM/IgG Test

2-8 degree celsius, 24 hours

Whole blood storage

2-8 degree celsius, 5 days

Plasma/Serum storage

20-25 minutes

Read results at

Primary dengue infection

IgM POSITIVE

Secondary or past dengue infection

IgG POSITIVE

Late primary or early secondary dengue infection

IgG and IgM POSITIVE

15-30°C

The temperature of the test area

2-30°C

The temperature during storage

60 grams per liter

ALBUMIN

20mg/dL

BILIRUBIN

442 micromol/L

CREATININE

3.4 μmol/L

EDTA

55 mmol/L

GLUCOSE

3,000 U/L

HEPARIN

4.34 mmol/L

SALICYLIC ACID

3.8%

SODIUM CITRATE

1,000 mg/dL

HUMAN IgG