Your SlideShare is downloading. ×
Immunologic laboratory-tests
Upcoming SlideShare
Loading in...5
×

Thanks for flagging this SlideShare!

Oops! An error has occurred.

×

Saving this for later?

Get the SlideShare app to save on your phone or tablet. Read anywhere, anytime - even offline.

Text the download link to your phone

Standard text messaging rates apply

Immunologic laboratory-tests

726
views

Published on

Published in: Technology

0 Comments
1 Like
Statistics
Notes
  • Be the first to comment

No Downloads
Views
Total Views
726
On Slideshare
0
From Embeds
0
Number of Embeds
0
Actions
Shares
0
Downloads
83
Comments
0
Likes
1
Embeds 0
No embeds

Report content
Flagged as inappropriate Flag as inappropriate
Flag as inappropriate

Select your reason for flagging this presentation as inappropriate.

Cancel
No notes for slide

Transcript

  • 1. Immunologic Laboratory Tests
  • 2. • Agglutination reactions• DAT• IAT• Immunofluorescence• ELISA• Western blot• Flow cytometryImmunologic Lab Tests Outline
  • 3. • What does it measure?• Where does the Ag-Ab interaction occur?• How is the Ag-Ab complex detected?Things to Remember About Each Test
  • 4. • Agglutination reactionsImmunologic Lab Tests Outline
  • 5. • Detection of Ag or Ab in patient specimen• Examples:• blood typing• testing for antibodies to infectious agents• testing for Hemophilus influenzae type B capsularantigen in CSFAgglutination Reactions: Purpose
  • 6. • Use particles coated with Ag or Ab• Add patient’s serum (containing Ab or Ag)• See if particles clumpAgglutination Reactions: Method
  • 7. Clumping = patient has the antibody (or antigen)Agglutination Reactions: Interpretation
  • 8. negative positive
  • 9. • Agglutination reactions• DATImmunologic Lab Tests Outline
  • 10. • Detection of Ab (or C’) on patient’s red cells• Also called the direct Coombs Test• Performed in patients with hemolytic anemiaDAT: Purpose
  • 11. • Use patient’s red cells (coated with Ab)• Add anti-human globulin (AHG) (Coombs reagent)• Look for agglutinationDAT: Method
  • 12. patient red cells + AHG = agglutination
  • 13. Clumping = patient red cells are coated withantibody and/or complementDAT: Interpretation
  • 14. • Agglutination reactions• DAT• IATImmunologic Lab Tests Outline
  • 15. • Detection of antibodies to red cell antigens• Also called the indirect Coombs Test• Performed as part of pre-transfusion testing• antibody screen• cross-matchIAT: Purpose
  • 16. • Use patient serum (containing Ab)• Add donor RBCs (coated with Ag)• Add anti-human globulin (Coombs reagent)• Look for agglutinationIAT: Method
  • 17. patientAb+ AHG = agglutinationdonorRBC=Ab-coateddonor RBC+
  • 18. patient serum(without red cell Ab)AHGreagent RBC(with red cell Ag)patient serum(with red cell Ab)AHGreagent RBC(with red cell Ag)ANTIBODY SCREENINGno agglutination(negative test)agglutination(positive test)
  • 19. Clumping = patient has an antibody to thedonor (or reagent) red cellsIAT: Interpretation
  • 20. • Agglutination reactions• DAT• IAT• ImmunofluorescenceImmunologic Lab Tests Outline
  • 21. • Detection of a specific antigen in a specimen• Examples:• detection of bacterial organisms• detection of antigen-antibody complexesImmunofluorescence: Purpose
  • 22. • Fix specimen on slide• Add antibody specific forthe desired antigen• Look for fluorescence• Fix specimen on slide• Add antibody specific forthe desired antigen• Add second antibody• Look for fluorescenceDirect IndirectImmunofluorescence: Methods
  • 23. Fluorescence = patient has the antigenImmunofluorescence: Interpretation
  • 24. • Agglutination reactions• DAT• IAT• Immunofluorescence• ELISAImmunologic Lab Tests Outline
  • 25. • Detection of antibodies in patient specimen• Examples:• home pregnancy tests• HIV tests• tests for some coagulation factors, cytokines,and autoantibodiesELISA: Purpose
  • 26. • Add patient specimen to well coated with ligand• Add AHG with enzyme attached• Add substrate• Measure color changeELISA: Method
  • 27. Color change = patient has the antibodyELISA: Interpretation
  • 28. Sandwich immunoassay• detects antigen (not antibody)• coat well with antibody• rest is like ELISARadioimmunoassay• detects antibody or antigen• detector is a radioactive substance• otherwise like ELISA or sandwich immunoassayELISA: Variations
  • 29. • Agglutination reactions• DAT• IAT• Immunofluorescence• ELISA• Western blotImmunologic Lab Tests Outline
  • 30. • Detection of antibodies in patient specimen• Most common example: HIV testWestern Blot: Purpose
  • 31. • Make a protein suspension of the target of theantibody you’re looking for (e.g., HIV)• Electrophorese the suspension onto a little gel strip• Apply the patient’s specimen (containing antibodies)to the strip• Add AHG that has an enzyme attached• Add substrate and look for bandsWestern Blot: Method
  • 32. Bands on strip = patient has antibodies tocorresponding proteinsWestern Blot: Interpretation
  • 33. Enough bands = patient is “positive”
  • 34. • Agglutination reactions• DAT• IAT• Immunofluorescence• ELISA• Western blot• Flow cytometryImmunologic Lab Tests Outline
  • 35. • Characterization of cell size, complexity, antigens• Examples:• diagnosis of leukemia and lymphoma• determination of CD4/CD8 counts in patientswith HIVFlow Cytometry: Purpose
  • 36. Complicated! Combine size, complexity andantigen expression data to come up withmeaningful description of cells.Flow Cytometry: Interpretation