1. Immunofluorescence Indirect Direct proof of antibodies (Ab)
proof of antigens (Ag)
antibody against Ab+ colour
Ab+ fluorescent colour (factory-made)
serum (IgG)
serum (Ag)
Ag (f-m)

2. Immunofluorescence
Task 1 and 2

3. ELISA I. (Enzyme-Linked Immunosorbent Assay)
Is used in a proof of antigens and antibodies

4. ELISA: proof of antigen
colour change + -
Substrate
Ab marked with enzyme
Serum with Ag
Serum without Ag
Ab bounded in well

5. ELISA II. – proof of antibodies
We detect classes of antibodies – IgM, IgA, IgG
!!! IgM and IgA demonstrate an acute infection
IgG generally = infection in past, but !!
exist so-called low avidity IgG – binding of these antibodies in reaction is not solid, it could be damaged by urea (they are present in earlier phase of infection)

6. ELISA: proof of antibodies - general+ -
Substrate
Ab against antibody marked with enzyme
Serum with Ab
Serum without Ab
Ag on the bottom

7. ELISA: proof of antibody classes – binding (capture)+ -
Substrate
Ab against antibody marked-with enzyme
Ag (factory-made)
Serum without Ab/with other Ab than searching
Serum with Ab (IgM/IgG)
Ab against IgM//IgG bounds all IgM/IgG in sample

8. Westernblott (proof of Ab)
Prepared Ag
gelELFO
parted
splitted by DDS Ag
Suck to cellulose membrane
Cutting
Strips

9. Westernblott Binding Ab to Ag + serum with antibodies (Ab)
OspC
+ serum with antibodies (Ab)
Firm-made strip with Ag
(BmpA, OspC, p39 etc.)
sample
pattern
At least 2 lines similary with pattern = presence of antibodies
Exception:ab against OspC u IgM line
ab against vlsE u IgG is enough

10. Pattern task no. 5 OspC is high specific
IgM against OspC present = acute infection, we don´t need to find IgM against other antigens