RC7990: In-Tray Exercise NAME:…………………..………………………………………………..

You have 20 minutes maximum to please answer the following questions:

1. Please explain what the acronym UKAS stands for and its role in pathology services.
2. You are setting up some manual assays for ant-inuclear antibody testing that require dilutions
1/80, 1/640 and 1/2560 to be performed. Please indicate how you would make a serial dilution
by filling in the blanks and using the pipettes and diluents available. The starting 1/80 dilution
would require a final volume of 400μl.

Pipettes Available Diluents Available

10 –100μl Single-Channel PBS + BSA
10–100μl 8-Channel Water
20 –200μl Single-Channel PBS
2 –20μl Single-Channel
100 –1000μl Single-Channel

………. μl 1/80 ………. μl 1/640

………. μl serum

………. μl diluent ………. μl diluent

………. μl diluent

1/640 1/2560
1/80

a. 1:80 dilution for HEp-2000 with a final volume of 400μl

Answer
Sample Pipette
Diluent Pipette
Diluent
Sample Volume
Diluent Volume

b. 1:640 dilution for serial dilution with a final volume of 400μl

Answer
Sample Pipette
Diluent Pipette
Diluent
1/80 Sample Volume
Diluent Volume

c. 1:2560 dilution for next serial dilution with a final volume of 400μl

Answer
Sample Pipette
Diluent Pipette
Diluent
1/640 Sample
Volume
Diluent Volume
3. Please can you explain using an example, the principle of indirect immunofluorescence? How is
this immunoassay used to aid in the diagnosis of immunological conditions? Where do you see
the future of immunofluorescence?
4. Listed below are a number of techniques used in an immunology laboratory, please match what
techniques are best used with the following disease associations.

Electrophoresis & Autoimmune Disorders

Immunofixation

Immunophenotyping Multiple Myeloma

Flow cytometry

Antigen-Antibody
Allergy & Total IgE
Detection by ELISA &
Indirect
Immunofluorescence Immunodeficiency &
Haematological
malignancies
Oligoclonal Band
Detection
Isoelectric Focusing
Pemphigus Vulgaris &
Bullous Pemphigoid
ISAC or IMMUNOCAP Blistering Skin Disorders

Biopsy Section
Acute Phase response
& Direct
Immunofluorescence

Immunochemical Proteins Multiple Sclerosis

Nephelometry or
Turbidity