METHOD FOR MODELING BIOFILMS FORMED BY VIBRIO CHOLERAE 01 SEROGROUP ON CHITIN SURFACE

摘要

FIELD: microbiology.;SUBSTANCE: invention relates to medical microbiology. Described is a method for simulating biofilms formed by Vibrio cholerae 01 serogroups on chitin surface. Method involves using chitin to form a biofilm formed by V. cholerae cells, followed by incubation and analysis of the results in PCR. Chitin used is Astacus astacus river cancer plates in amount of 10–15 pieces and size of 0.3×0.3 cm, which are placed into a bottle with capacity of 100 ml, containing 30 ml of tailed river water. Bottle is autoclaved for 10 minutes at 120 °C, and after cooling, a suspension of Vibrio cholerae 01 serogroup cultures is introduced into the bottle to a final concentration of 10⁴ m.c./ml. Incubation is performed at 10 °C and 28 °C. Analysis of the results of formation of both simple and complex V. cholerae biofilms is carried out in PCR using primers to INDEL-locus 1699: VC1699-1 GCTTAGCTATTTTTGGGTATAGGTT, VC1699-2 CGTTCATTTTTACTCAAACAGTCA. Atoxigenic strains of ctx-tcpA- form an amplicon with mass of 132 base pairs, and toxigenic strains ctx+tcpA+ form an amplicon with weight of 116 base pairs.;EFFECT: thus confirming the presence of toxigenic strain cells in a complex biofilm and the ability to colonize the surface of chitin.;4 cl, 3 dwg, 4 ex