Techniques in Immunology

Techniques in Immunology are divided into 2 major categories 1- Humoral 2- Cellular Humoral technique: is used to measure o Anti-body o complement o acute phase protein except C-reactive protein o soluble mediators of immune response For example, if I suspected a disease that affects the complement system, I can measure C2, C4...Etc.
 Humoral Tech are divided into 4 •

1- Precipitation reaction  It is based on antigen-antibody reaction in Solution.  Both antigen & antibody are soluble.  It is the simplest, easiest & least sensitive.  It is based on precipitation curve  If I have fixed amount of AB and I keep increasing the amount of AG with I will form immune complex which will precipitate.  We have 3 zones: First  zone of AB excess. Middle zone of equilibrium. Last zone of AG excess.
o A kind of precipitation reaction is double gel diffusion, which is

a precipitation reaction that takes place in gel like the one we saw in lab on agars. It is used in identifying relationship between different antigens.
o RID (radial immuno-diffusion) It is used in measuring

Immunoglobulin concentration in serum; Quantitaion of AG or AB.
o Immuno electrophoresis used to study gamma region. To

diagnose hypo or hyper-gamma globulinemia or multiple myloma. I expect from immune electrophoresis presence or absence of the following: monoclonal band, class of immunoglobulin, type of light chain.
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2- Agglutination reaction  It is antigen-antibody reaction BUT  Either AB or AG are INSOLUBLE  More early, more widely used than precipitation, and is more sensitive.  Examples of clinical uses of agglutination: • RPR ( rapid plasma reaging) for screenings for syphilis • Diagnosis of mycoplasma pneumonia(old agglutination) • Diagnosis of certain bacterial infections e.g. prosilosis • Measure Rheumatoid factor • ASO (anti-streptolysin) • CRP( c-reactive proteinacute phase protein)
3- Labeled immunoassay these include:

 Florescence immunoassay (FIA)  Radio immunoassay (RIA)  Enzyme immunoassay (EIA) (For details ... refer to the original tafree'3 ... this is just an outline) 4- Compliment fixation assay  Antibody + antigen=activation of complement. The indicator system is usually sheep RBC.
 Cellular technique: to measure the

o Number of cells of immune system. o Function of cells of immune system.

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