FIELD: medicine; microbiology.
SUBSTANCE: method an be used for specific identification and evaluation of potential ability of non-cultivated forms of vibrio cholerae O1 and O139 serous groups to reversion, which vibrios take place in water solutions. Method is based upon preliminary preparation of bacterial suspension of non-cultivated forms of vibrio cholerae O1 and O139 serous groups and bacterial suspension of reference culture of Vibrio Cholerae O1 and O139 serous groups. The working dilution of bacterial suspensions and monoclonal antibodies conjugate is determined and specific monoclonal antibodies are selected for conservative and labile lactose-peptone medium epitopes followed by dot-immune-enzyme analysis or solid phase immune-enzyme analysis reaction. Then appearance of color point or optical density, being higher than 0,2-0,65, is registered and conclusion is made on presence of non-cultivated forms of Vibrio Cholerae O1 and O139 serous groups in sample and their potential reversion ability. Preliminary preparation of non-cultivated forms is performed due to increase in their concentration up to 106-107 mxkl/ml followed by inactivation. For the purpose 20 mkl of 4% sterile gentamicine solution is added to 1 ml of microbe suspension followed by subsequent cooling of samples down to 3-4 deg C during 2 hours. Bacterial suspension of control culture Vibrio Cholerae O1 or Vibrio Cholerae O31 is made from 18-hours culture, grown onto Martin's agar. After that the suspension is used for preparation of bacterial suspension in physiological solution, pH 7,6 till final concentration of 107 mxkl/ml. Method uses set of specific monoclonal antibodies, got to different epitopes of O-antigen of lactose-peptone medium cultures of Vibrio Cholerae O1 and O139 serous groups: 3A9, D11, 3E4, 4H5, D8D6, C3B4, F8G12, F11E5.
EFFECT: ability of getting express-answer at low concentration of tested cells.
5 cl, 2 tbl
