A monochromatic wavelength is introduced to the wells containing antigen and antibody
complex, which result in absorbance and reflection through the protein. Also, some additive
could be used for fluorescence and luminescence.
 Slide 4: Provides a pictorial representation of working of an ELISA plate in terms od
antigen antibody interaction
In the case of sandwich ELISA, pair of antibodies are used, in which one antibody act as a
capture antibody and the second antibody work as a detection antibody which could either be
monoclonal or polyclonal antibody in nature.
The highly selective nature of antibody results in the capturing of antigen’s epitope.
The Sandwich ELISA test is considered more effective for the antigen with two epitopes.
In a typical sandwich, ELISA test following steps are followed
Coating of ELISA plate well with the capture antibody
Addition of antigen (sample)
To this Antibody antigen mixture, a detector antibody is added, which generally has an enzyme
attached to it and subsequently incubated for a fixed interval of time.
Followed by the addition of chromogenic substrate and the concentration are measured with
ELISA plate reader.
 Slide 5: Discuss regarding the application of ELISA plate reader in the detection of
diseases
Diseases detection using micro plate reader
The Micro plate Readers have been used for the detection of many diseases such as
▪ Dengue: Fever occurs due to dengue virus, detection is carried out based on the antigen
related to the virus.
▪ The Virus mode of transmittance from one person to other is through mosquitoes.
▪ Major problem for tropical and sub-tropical region.

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