1. Mantoux – in vivo detection of specific cell immunity after exposition to antigen - burden of patient by antigen - possible immunodeficiency of patients (anergy, risk of alergic reaction) - memory cell after BCG vaccination, - exposition to antigen = activation of MC - interpretation, - booster dose of antigen,

2. Matoux skin test reading application positive negative 15 mm

3. Measurement of induration 0 – 15 mm – negative, not enought, vaccination, revaccination 15 mm – OK, positive (after vaccination ?) 15 and more – too much, possible activity

4. in vitro detection of specific cell immunity after exposition to the antigen whole blood of patient to 3 vials covered with specific antigen (M. tbc) specific memory cells after disease are activated antigen is not given to patient in vivo - no risk of exposition of immunocompromised patients

5. Test based on exposition of whole blood in 3 vials - antigens of M.tbc, - 1 vial - non specific mitogen – positive control - without antigen – negative control

6. Vials are incubated 16 – 24 hrs at 37 dg C (sensibilised memory cells are exposed to specific antigen and start production of INTERFERONU GAMA – cytokín of CMI Centrifugation Harvest the plasma Storage

7. To detect IF-  secreted after in vitro activation (antigen on the surface of vial + memory cells in the whole blood of pacient) of memory cells produced after the infection (by M.tbc) we use ELISA test

9. Interpretation: comparison of concentration of IF  in the vial with -TB ag : negat.contr. Negat control – exclude the nonspecific reaction Pozit.control – demonstrate capacity to react (negat. in imunocompromised or annergic

10. comparison Mantoux: in vivo reactivity of memory cells produced after vaccination (BCG) and after disease (M.tbc) application of i.d. memory cells migrate to the place of injection inflamation with induration Quantiferon: in vitro memory cells after M.tbc infection present in the blood in vial + M.tbc antigen = release of IF  ELISA for detection

11. detection of cytokines In vitro ( immunity of patient is not influenced, boostered) specificity sensitivity not cross reacting antibodies PERSPECTIVES detection of latent infection (any infection if the antigen is available) – in practice now only M.tbc CMI - scientifical purposes

12. Other use Detection of latent infection CMV – in transplantation, tissue parasitoses – withou serological reaction, systemic mycoses, detection of CMI or memory cells – for identification of memory after vaccination, when antibodies are no more present, detection of non responders