Immunological and Molecular Diagnositics
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What are the uses of serology?
To see if an animal has been exposed to a pahogen To test whether there has been a response to vaccination To test for immune-mediated disease
What can immunoassays be used for?
Looking doe a specific pathogen in a sample Measuring biomarkers Immunophenotyping
When taking a blood sample, how does the collection method differ when wanting: a) Serum b) Cells
a) Clotted sample b) Anticoagulant (citrate/heparin)
What is the serological marker for innate immunity?
Acute phase proteins
What do measurements of the following show: a) Total IG b) Antigen specific IG?
a) Detecting failure of passive transfer Ig deficiency syndromes Gammopahies b) Exosure o specific pathogen Response to vaccination Dx of antibody mediated hypersensitivity
In a recent infection what should one test for if using an ELISA?
Antigen - as an adaptice immune response may not have yet developed.
If antibodies are seen in response to infection, how can the progress of infection be determined?
Look at the type of Ig produced and take samples 2-3 weeks apart.
Which type of ELISA is used to detect: a) Antigen b) Anibody
a) Sandwich b) Indirect
Describe a VN assay and why is this useful?
Infect cells without serum to see positive result and determine whether virus has a cytopathic effect - otherwise will need to visualise with IFA. Then add serial dilutions of serum and see the titre that prevents cells being infected. This is advantagous c.f. ELISA/IFA as it indicates that antibodies are biologically active.
How can T cell responses be evaluated in vivo and in vitro?
In vivo - Intradermal skin test (leave >72hrs for delayed type hypersensitivities) In vitro - measure cytokine release following Ag stimulation. (may also be able to look at surface receptors and proliferation??)
What other things can be tested to test immune funcion?
• Immunodeficiency - Neurophil function - if defective adaptive responses • Allergy e.g. Allercept testing • Autoimmune disease - specific assays for autoantibodies are available
How can a pathogen be detected in a sample?
Sandwich ELISA IFA IHC
What methods are there for measuring biomarkers (e.g. hormones) in a sample?
Chemiluminescence RIA
What is immunophenotyping?
Using antibodies against cell surface markers to determine cell types within a sample.
When testing an animal for genetic disease what should be considered?
What type of mutation (insertion/deletion/substitution) Is disease simple or complex genetic disorder Is it autosomal or sex linked Dominant or recessive Complete or incomplete penetrance (i.e. does presence of the mutation always = disease?
What is DNA genotyping useful for?
Establishing parentage Diagnosis of disease Screening for carrier animals Establishing risk of disease in later life