STUDY OF EXTRACELLULAR ANTIGENS BYIMMUNODIFFUSION METHODS IN DIFFERENTIATIONOF PATHOGENIC BURKHOLDERIAE

Aim. Isolation and composition comparison
of extracellular antigens (ECA) of pathogenic
burkholderiae in SDS-PAGE electrophoresis
and their use for differentiation of these microorganisms
by immunodiffusion methods.
Materials and methods. 60 Burkholderia pseudomallei
strains, 14 B. mallei strains, 5 B. thailandensis
strains, 4 B. cepacia strains were
studied. ECA was obtained by Liu technique on
F-agar covered with cellophane. SDS-PAGE
electrophoresis was performed in 10% gel by
Laemmli, immunodiffusion reaction (IDR) in
1% agarose gel, IDR with live cultures, immunoelectrophoresis
(IEPH) was performed by the
standard techniques. Sera was obtained by immunizing
rabbits with a mixture of ECA and
incomplete Freund adjuvant. Results. ECA
spectra of typical strains of the studied burkholderiae
strains after the electrophoresis in SDSPAGE
stained by silver have 8 - 9 major fractions.
ECA electrophoregrams of B. pseudo -
mallei and B. thailandensis had a high simi larity.
ECA analysis by IDR with antisera against ECA
revealed maximum number of cross-reactive
ECA (3) between B. pseudomallei и B. thailandensis.
These strains had only a single crossreactive
ECA to B. mallei strain. IDR with live
culture and antisera to B. thailandensis ECA
revealed ECA in all the B. pseudomallei, B.
thailandensis strains and did not reveal those in
B. mallei strains. Analysis of electrophoregram
obtained with IEPH method of pathogenic
burkholderiae ECA with antisera to ECA revealed
differences of the composition sufficient
for their differentiation. Conclusion. The differences
of ECA composition revealed by immunodiffusion
methods allowed to develop additional
approaches of differentiation of glanders
and melioidosis pathogenic agents.

Burkholderia pseudomallei, B. mallei, B. thailandensis, pathogenic bacteria, Burkholderia pseudomallei, B. mallei, B. thailandensis, extracellular antigens, immunodiffusion methods