8. Clinical immunology Flashcards
2 types of immunodiffusion
- radial
2. double
8 types of immunoassays
- radio
- ELISA
- flow cytometry
- multiplex
- microarrays
- complement
- immunofluorescence
- immunocytochemistry
difference between radial and double immunodiffusion
radial - 1 well
double - many wells
what occurs in immunodiffusion?
Ag and Ab form complexes - forms ring of precipitate
radial ID diagnoses .. (2)
- dysfibrinogenaemia
2. brucellosis
what is dysfibrinogenaemia
coagulation disorder
what is brucellosis
bacterial infection from food
Double ID diagnoses.. (3)
- fungal inf
- viral inf
- forensics
other than ID, which other lab method uses precipitation?
Agglutination
4 uses of agglutination testing
- blood group serology
- Thyroid testing
- Rose Waaler test - rheumatoid arthritis
- Paul Bunnel test - glandular fever
what is immunoelectrophoresis?
Biochemical method for separating proteins
what 2 processes does immunoelectrophoresis use?
electrophoresis and immunodiffusion
Describe steps in immunoelectrophoresis
- sample added to gel
- electric current applies
- Ig migrate through gel
- antisera added
- Ag/Ab complex visualised
what is immunoelectrophoresis replaced by? 3 reasons why
immunofixation
- easier to interpret results/perform
- faster
- more sensitive
difference between immunoelectrophoresis and immunofixaton
IF uses an acid violent stain for interpretting
what does immunoelectrophoresis and immunofixaton diagnose? - 4
- Waldenstrom’s macroglobinaemia
- myeloma
- Ig type
- allergy testing
what is Waldenstrom’s macroglobinaemia ?
high level of IgM
which process is used to commercially produce Ab?
immunoassays
what are immmunoassays generally used to measure concentrations of? (4)
- hormones
- tumour markers
- autoAb
- allergens
3 types of marker are attached in immunoassays, what are they?
- enzyme; HRP, alkaline peroxidase
- Radioisotopes - I-125, H-3, P-32
- fluoresents - fluoresein, rhodamine
why are radioisotopes not usually used?
health and safety
in ELISA - which 2 ways are used to measure colour?
- optical density
2. absorbance
in ELISA- what is the intensity of colour proportional to?
Amount of Ag bound
what 3 things can flow cytometry identify?
- leucocytes
- immunophenotyping (diseases)
- allergy testing
in flow cytometry - what will be identified in those with a latex allergy?
CD64
in flow cytometry - what will be identified in those with chronic lymphocyte leukaemia ?
CD5
in flow cytometry - what will be identified in those with HIV?
CD4
which method uses polymerase chain reactions?
multiplex technology
describe process of luminex - multiplex technology
- polystyrene beads have Ag and fluorescent dyes attaches
- , Beads attach to Ag in serum
- Analysed in flow cytometer
name 5 things multiplex technology detects?
- Ig isotypes
- viral resp infections
- virus causing eye infections
- rheumatoid arthritis
- cytokines
- Growth factors
- acute phase proteins
- Ab
which method tests for breast cancer? what does it test for specifically:?
Microarray
BRCA1 and BRCA2
which test is commonly used for allergy testing
microarray
describe steps in microarray allergy testing
- common allergen proteins attached to glass
- Pt plasma added
- Ag/Ab reaction
- Bound IgE detected
- microarray scanner determines amount of fluorescence
4 methods used for allergy testing
- skin prick test
- patch test
- lab tests
- challenge test
what does immunofluorescence detect?
antinuclear Ab- bind to contents of a cell
in what type of diseases are there high levels of antinuclear Ab?
autoimmune
describe set-up in immunofluorescence reactions
- slide w/Ag
- Ab attached from pt
- secondary Ab attached which is conjugated to fluorescein
name 5 diseases immunofluorescence can detect
- Autoimmune (AI) liver D
- AI kidney D
- AI thyroid D
- coeliac D
- Diabetes
- Addison’s D
- Pernicious anaemia
which methods are used to detect tumours - adenocarcinoma, lymphoma, breast cancer, ovarian cancer?
immunocytochemistry, immunohistrochemistry
