#11 Immunological methods and Techniques

Question 1

What does precipitation measure?

Answer 1

The amt of immunoglobulins (IgM, IgG, etc) in blood

Question 2

What is direct assay (DAT) used for?

Answer 2

Used to test for autoimmune hemolytic anemia (low RBC count)

To detect RBC Ags or Abs against RBC Ags

Used when hemolytic anemia is suspected, Rh incompatibility, ABO blood grouping

Draw blood, wash away unbound Ab, incubate in anti-human globulin which binds to patients Ab, resulting in agglutination.

+ result: suggests immune hemolytic anemia (because we know that Ab are binding RBCs)

Question 3

What is indirect assay (IAT) used for?

Answer 3

Used in prenatal testing of pregnant women and in testing blood prior to a blood transfusion. It detects antibodies against RBCs that are present unbound in the patient’s serum

Only deals with plasma (unlike DAT which still has RBC, we get rid of RBC and just want the Ab). RBC’s are added that are used to detect certain Ab. Binding occurs, then add anti-human globulin results in agglutination

+ result: indicates presence of RBC antibody in the patient

Question 4

CD markers for flow cytometry

Answer 4

All leukocytes: CD45
Monocytes: CD14
T reg: CD25
NK cells: CD56