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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

    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

    Dengue Test (IgG & IgM)

    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.

    IgG & IgM Elisa
    Dengue Elisa

    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.

    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

    Possible References Used

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