Dengue Test Procedure

Test Procedure:

There are two methods are explained

ICT (Screening) Method

ELISA Method

ICT Method

ICT (Immuno chromatographic TEST) is a initial test method its called Screening Method.

Test Requirements

• Patient’s Sample
• Centrifuge
• Test Tubes
• Micropipettes
• ICT Device
• ICT Device Buffer
• Yellow Tips
• Patient’s Performa for Report

Some Laboratories use Dropper and they not use micropipettes.

Reagent Prepration

ICT Device Buffer is ready to use and it is room temperature item.

Test Procedure

• Open the ICT Device and mark it as pt. name or number
• add 10ul Patient’s Serum.Plasma
• Add 2 drops ICT Device Buffer
• Wait for 5-10 mints
• Read The Result

Some ICT Device companies methods are different. That is why read the manual carefully.

Result Preprations

See the below chart

ELISA Method

ELISA (enzyme-linked immunosorbent assay) is a plate-based assay technique designed for detecting and quantifying substances such as peptides, proteins, antibodies and hormones. Other names, such as enzyme immunoassay (EIA), are also used to describe the same technology.

Test Requirements

There are Basic Requirements for this Technique.

Patient’s Sample

Incubator

Centrifuge

Micropipettes (10ul – 100ul & 100ul – 1000ul)

Microplate Washer or Manually Washing Bottles

Test Kits (EIA)

Plate Reader

Patient’s Performa for Report

Contents of the test kit:

Microplate wells (12×8)

Calibrator (1×2.0ml)

Positive control (1×2.0ml)

Negativecontrol (1×2.0ml)

Enzyme conjugate (1x12ml)

Sample buffer (1x100ml)

Wash buffer (1x100ml)

Chromogen/substrate solution (1x12ml)

Stop solution (1x12ml)

Test Instruction (1 booklet)

Reagent Prepration

All Reagents and samples temperature must 37°C

Patient’s Sample:

The patient samples for analysis are diluted 1:101 with sample buffer. For example, add 10 µl serum to 1.0 ml sample buffer and mix well. Incubate the mixture for at least 10 minutes at room temperature.

Wash Buffer:

The wash buffer is a 10x concentrate (1 part reagent plus 9 parts distilled water).

Test Procedure

Add 100ul Standards, NC, PC and Diluted Samples in each well(See Reagents Preprations)

Incubate 30 mint in Room Temperature

Wash Wells 3 times with 400ul Wash Buffer(See Reagents Preparation)

add 100ul Conjugate in each Wells

Incubate 15 mint in room temperature

Wash Wells 3 times with 400ul Wash Buffer(See Reagents Preparation)

Add 100ul Substrate/TMB/chromogen Solution in each well

Incubate 20 mint at room temperature

Add 50 Micro Stop Solution in each well

colour intensity should be made at a wavelength of 450nm and a reference wavelength of between 620nm and 650nm within 30 minutes of adding the stop solution.

Result Preprations

This Procedure is same of 80% of Different Companies kits.

Cut-off:
Standare Value = 0.5 (some companies are different)
Std abs = 0.127
Cut-off = std abs + std value
= 0.127 + 0.5 = 0.627
Patient Result:
Pt abs = 0.243 = Non Reactive
Pt abs = 0.634 = Borderline
Pt abs = 0.843 = Reactive

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Possible References Used

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