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,
Matoux skin test reading
application positive negative
15 mm
Measurement of induration 0 – 15 mm – negative, not enought,
vaccination, revaccination 15 mm – OK, positive (after vaccination ?) 15 and more – too much, possible activity
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
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
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
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
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
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
detection of cytokines In vitro ( immunity of patient is not influenced, boostered) specificity sensitivity not cross reacting antibodiesPERSPECTIVES detection of latent infection (any
infection if the antigen is available) – in practice now only M.tbc CMI
- scientifical purposes
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