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محمود روكانA4 احمد محمدA1 اسل عدنانA1 PDF

1. The document discusses radioimmunoassay (RIA), an in vitro assay that can detect antigens with high sensitivity using competition between labeled and unlabeled antigens binding to antibodies. 2. RIA can be used to detect viral, bacterial, and other infections by measuring hormones, vitamins, drugs, and other biological substances. 3. The procedure involves adding labeled antigen to wells coated with antibody, then adding the unlabeled antigen from a sample to compete for binding. The level of radioactivity measured after washing indicates the concentration of antigen in the sample.

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59 views6 pages

محمود روكانA4 احمد محمدA1 اسل عدنانA1 PDF

1. The document discusses radioimmunoassay (RIA), an in vitro assay that can detect antigens with high sensitivity using competition between labeled and unlabeled antigens binding to antibodies. 2. RIA can be used to detect viral, bacterial, and other infections by measuring hormones, vitamins, drugs, and other biological substances. 3. The procedure involves adding labeled antigen to wells coated with antibody, then adding the unlabeled antigen from a sample to compete for binding. The level of radioactivity measured after washing indicates the concentration of antigen in the sample.

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AL-KITAB UNIVERSTY

College of Technology Medical


Department of medical Analysis
Stage(3)

Viruses
(Radioimmunoassay)

Supervision :DR.wirya

Preparation:
1-Mahmoud rokan /A4 2- Ahmed mohammed/A1
3- asal adnan/A1
Introduction
Radioimmunoassay (RIA) is an in vitro assay that measures
the presence of an antigen with very high sensitivity
depend on competition reaction between two types of
antigen ( labeled( with radioactive substance) + unlabeled
antigen). Basically any biological substance for which a
specific antibody exists can be measured, even in Minute
concentration

Types :
1 - Direct
2 – Indirect
For doing the test three main thing shall be considered

include
1- Anti antibody for the Ag interest ( adsorbed in microtiter
wells)
2- 2- Labeled Ag with I125 ( Hot Ag) from kit.

3-Unlabeled Ag ( cold Ag) from sample.

The test is used for detecting:


1- Viral infection.

2-Bacterial infection.

3-Hormones.

4-Vitamiens.

5-Drug.
REQUIREMENTS
Micro titer plate / Test tubes.
Pure antigen
Radio labeled antigen
Antibodies
Standards
Centrifuge
Radioactive counter
Procedures
1-We add the labeled Ag ( Hot Ag) to the microtiter that
adsorbed with Anti A antigen ( Ag of sample ( serum) )

2-Washing to remove unbound Ag

3-The radioactivity in this time is 100%.

4-Then we add the sample Ag ( A antigen( cold Ag) in


minimum amount ( in Nano Gram).

5-The cold Antigen occupied the hot Antigen place and


competition reaction occurs between two types of Antigens.

6-Washing to remove unbound cold antigen

7- The radioactivity change in this time ( not still 100%)


depend on the concentration of cold antigen.

8- Then we measure the radioactivity and draw the curve to


know the concentration of Antigen( cold antigen) in the
sample.
Advantages
Highly specific: Immune reactions are specific, the greater the
specificity of the antiserum, the greater the specificity of the
assay.
High sensitivity Immune reactions are sensitive, Using
antibodies of high affinity it is possible to detect a few
picograms (10-12 g) of antigen in the tube. » Accuracy and
Precision

Disadvantages

Radiation hazards: Uses radio labelled reagents


Requires specially trained persons
Labs require special license to handle radioactive material
Requires special arrangements for
 Requisition, storage of radioactive material
 radioactive waste disposal

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