ELISA is defined as

an “immunological” test that uses

enzyme linked antiglobulin & substrate
bound to an “inert surface”.

Assay-A quantitative or qualitative test of a substance (especially an ore or a drug) to determine its components; frequently
used to test for the presence or concentration of infectious agents or antibodies etc.
 ELISA
The most important Serological Test

ELISA is a “Serological Test” which is used for either detection of

Ag or Ab.
A reaction is said to be “Serological”; when Ag-Ab reactions are
carried out in-vitro.
This is a
Serological Test.
See the in-vitro
Ag-Ab reaction
being carried out.

This is ELISA plate.

See the 96 walls on
the plate with
12X8 arrangement
Why ELISA is so important??????
ELISA is a sensitive laboratory method used to detect the
presence of antigens (Ag) or antibodies (Ab) with
a wide variety of biological samples.

In these kind of reactions , an absorbing material specific

for one of the components i. e. Ag or Ab which are used.

ELISA is many times referred as “Solid-Phase Immuosorbent

Assay” (SPIA) by some professionals due to the fact that
Ag or Ab are coated to solid phase.
Ag or Ab is first coated to TT wall. To this respective Ab or Ag
Is added. To it then enzyme labeled molecule is added. When
substrate is added ; if reaction is positive then colour reaction
is quickly seen.
This is the generalized principle applied in all kinds of ELISA
Tests.
This is also called as SANDWICH ELISA.
The samples that are added are sandwiched in the layers of Ag & Ig
so it is called so.
LISA FOR DETECTION OF Ag IN FECES

Ab Secondary Ab Colour on adding substrate

Ag Enzyme

1.Microassay plate coated with Ab.

2.Ag collected in sample gets coated over Ab.
3.Adding Enzyme labelled Ab ; forming complex.
4.Adding substrate.
5.This forms coloured substrate i.e. test is positive. Substrate
splits & colouration is seen.
6.If test is negative, no coloured substrate is formed.
LISA FOR DETECTION OF Ab IN SERUM

Ag Ig-Ab with Enzyme

Ab Colour on adding substrate

1.Microassay plate coated with Ag

2.Ab in collected sample gets coated over Ag previously
present on micro assay.
3.Adding enzyme labelled Ag (here it may be IgG)
4.Adding substrate.
5.This forms coloured substrate indicating positive test.
Interpreting the result….

Positive Negative
(Green colour is seen) (No colour is seen)
loser view of ELISA PLATE

NEGATIVE INTERMEDIATE POSITIVE

Types of ELISA
ELISA is the widely used test in the “Clinical Serology”. So now a day ample
research work has been done. Newer techniques for increasing demands
of market are being satisfied by various developed & convenient types of
ELISA. These are:

1.CARD & DIPSTICK ELISA :

Simple diagnostic technique for bedside application.
Suitable for clinical laboratory.

2.CYLINDER OR CASSETTE ELISA :

The most rapid technique of ELISA that completes within 10 minutes.
To our surprise normal ELISA takes 2-4 hours.

3.SANDWICH ELISA :
Best technique for detection of circulating virus in blood dogs & cats.
APPLICATIONS OF ELISA-1

1 ELISA is used in detection of FPT.

FPT is “Failure of Passive Transport”.
FPT is failure of intestinal absorption of IgG from colostrum
which is required for protection of young one against enteric diseases.
Especially “Dipstick ELISA” is used here.

2 ELISA is a test used to detect and identify antiviral Ab of wide variety.

Here “Indirect ELISA” is used. This is the “simplest technique” available
in commercial market.
APPLICATIONS OF ELISA-2
3 ELISA is also used in detection of “food allergies”.
No doubt this of limited usefulness.

4 ELISA is also used for detection of Autoimmune Thyroiditis.

ELISA is used here to detect Ab against thyroid microsomal or colloid Ag.
Of or relating to microsomes
A tiny granule in the cytoplasm that is where
protein synthesis takes place under the direction of mRNA

5 ELISA is used to detect FeLV viremia. Feline Leukemia Virus

Here , whole blood sample is used.
An alternative technique is the use of saliva for test.
Even tears are also used.
Why only ELISA???

1. ELISA makes us enable to test widest variety of biological

samples to test at higher level of accuracy.

2. ELISA is safest as there is no radioactive material involved as in case of

radioactive assays which were used prior to ELISA at larger scale.

3. Recent developments in field of Applied Serology makes us to carry out

tests at commercial level as well.
REFERENCES:
2.Immunology & Serology by Tizzard.
3.Microbiology by Panikar
4.www.medscape.com
5.www.serology.com
5.For images www.google.com & www.yahoo.com

THANK YOU….
Sarang S. Desai.
V/05/0170.
GUIDED BY---
Dr. Mhase.
Department of Microbiolo