SU597235A1 - Method of producing erythrocytic plague diagnosticum - Google Patents

Description

The invention relates to medical microbiology, in particular, to methods for obtaining erythrocyte diagnostics for indication, identification of pathogens of the plague microbe and serological diagnosis of the diseases caused by them.

A known method of preparing an antigen erythrocyte diagnosticum, for which Baker fraction 1 (capsular antigen) is used as sensitin for erythrocytes without its prior activation.

However, using this diagnosticum only capsular (typical) strains of the plague microbe can be identified.

The aim of the invention is to obtain an erythrocyte diagnosticum, which makes it possible to identify both capsular (typical) and capsuleless

Salt (atypical) strains of plague microbz.

This goal is achieved by using the fraction V of the plague microbe as sensitin for erythrocytes.

The procedure for isolating the V fraction and preparation. On its basis, the erythrocyte diagnosticum is as follows;

The fraction Y is isolated from the capsule-free and toxic version of the EB when it is used (and increased concentrations of anionic surfactant sodium deoxycholate (DOX). After 4-fold exposure to low concentrations of DOX (from 0.00 to 0.32) on the bacterial mass and the subsequent removal of the supernatants, the residue is added to DOX to 1.285. 5 volumes of acetone are added to the supernatant, the precipitate is dialyzed and lyophilized.

Thus, fraction V is used as sensitin for erythrocytes.

To obtain a diagnosticum, formalinized sheep erythrocytes are taken, which are treated with a solution of Nina at a dilution of 1: l600QO for 13 min at 37 ° C. Then the erythrocyte is precipitated by centrifugation for 3 min at 2UU rpm. The erythrocyte sediment is washed three times with 20-30 times the amount of physiological solution pH-° 7, and adjusted to 2, the concentration with the same solution. The necessary amount of fraction V is diluted with saline: To a concentration of 1000 µg / ml and add DOX with this calculation so that its final concentration is%. After 13–20 min of contact at room temperature, a mixture of fraction V and DOX is added to red blood cells at the rate of 10 µg per 1 ml of 2.3–2 tanned red blood cells. After shaking, the roCites are placed in a fridge at: ib-lii h. For 2 hours before the end of the sensation procedure, erythrocytes add formalin before //. Then the erythrocytes precipitate; g .0T iT, T;: -: ((V, - ;; p); ai; |, lg; "; saDOK

washed three times with large amounts of normal rabbit serum diluted 1: 230 with saline. The erythrocyte sediment is adjusted to 10 with a concentration of a sucrose-gelatin medium (3 gelatosis + /, j% sahe roses on distilled water), poured into ml ampoules or penicillin vials and subjected to freeze drying in any vacuum drying apparatus. Residual moisture should not exceed more than 3% with ampoules and vials of dried red blood cells

sealed or hermetically sealed. The diagnosticum prepared in this way can be grmchit at room temperature.

The proposed method provides

0 identifying both cag, sulnyl (typical), and capsule-free (agipichny) atoxic strains of the plague microbe. This allows using it with great confidence in practice.

5 epizootological examination of natural foci of plague, as well as for identification and identification of typical and antigenically defective strains

: H- MANY M.iKO- / Ji;

Claims (1)

METHOD FOR PRODUCING ERYTHROCITAR DIAGNOSTICUM FOR PLAGUE by treating red blood cells with a formalin solution and then tannin, followed by sedimentation of red blood cells from the solution and their sensitization and isolation of the target product, characterized in that, in order to identify capsular and capsuleless microbial, plague, microorganism plague microbe,