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)

Immunofluorescence Task 1 and 2

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

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

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)

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

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

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

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 1 line ab against vlsE u IgG is enough

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