DENGUE FEVER TEST

Arranged By : 
Aisyah Hakim
Cesa Navratilovi
Elis Khopipah
Imron Adhitya Putra
Retno Aningsih
Risma Eka Damayanti
Roudhotul Fauziah
Yolan Aulia Putri

MEDICAL LABORATORY TECHNOLOGYST

POLYTECHNIC OF HEALTH
MINISTRY OF HEALTH
BANTEN
2018/2019
 DENGUE FEVER TEST
USING THE ELISA METHOD
 Brief purpose and application

IgG DHF contained in the sample will be related to anti

human IgG placed in the sample well. Then added HRP
monoclonal antibody (Mab) conjugate so that complex
compound are formed and release peroxide which
reacts with chromogen to form blue compounds whose
intensity is proportional to concentration of IgG DHF in
the sample. The reaction is stopped by adding sulfuric
acid as a stop solution so that the color changes to
yellow which the absorbance reads with an Elisa plate
reader at 450 nm and 620 nm.
Symptoms of people affected by DHF

1. Sudden high fever, for no apparent reason, persists

for 2-7 days
2. Sign of bleeding
3. Liver enlargement
4. Trombocytopenia (150.000 or less)
5. Hemoconcentration : hematocytes increas by 20% or
more compared to the value during treatment
Tool and Materials
1. Incubator
2. Elisa plate reader
3. Micropippete 1000 ml, 500 ml , 100 ml, 50 ml, dan 5
ml.
4. Rack well and it’s cover
5. IgG DHF reagent kit
6. Yellow tip
7. Washing solution
8. Sample
9. Aquadest
10. TMB
 Flow Chart

Pippete 1000 µL diluent

Add 10 µL serum sample
Homogeneous
Pippete 100 µL from mixture
Incubation
Wash with washing solution
Add 100 µL conjugat enzym IgG
incubation
Add 100 µL TMB
incubation
Add 100 µL H2SO4 2N
Read with elisa reader
 Detail Procedure
1. pippete 1000 µL diluent serum and 10 µL serum samples,
Negative Control, Positive control, calibrate into a test tube.
2. Shake until homogeneous
3. In the 100 µL pippete the mixture is inserted into each well.
4. Incubated for 30 minute at 37°C
5. Washed each with 6 times the washing solution
6. Add 100 µL conjugat enzym IgG
7. Incubated for 30 minute at 37°C,than Washed each with 6
times the washing solution
8. 100 µL TMB added to each well
9. Incubated for 10 minutes at room temperature
10.Added 100 µL H2SO4 2N
11.Read absorbance with Elisa Reader
Reading and interpretation
N Ratio Result Interpretation
o
1. IgG< 1,8 Negativ No secondary
e infection
2. IgG 1,8- Equivok Need to retest
2,2 al
3. IgG > Positive Suspected active
2,2 secondary
infection
 Calculation
Cut off (CO) : Absorbance calibration x factor (0,62).
Negative : if absorbance < cut off
Positive : if absorbance > cut off
 O U
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N K
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