SU1726510A1 - Strain of bacteria vibrio nag 083 serovara, used for preparation of monovar diagnostic serum - Google Patents

Abstract

Use: medicine, diagnostics, microbiologists - obtaining serum to identify pathogenic vibrioes. SUMMARY OF THE INVENTION: A culture of a Vibrio Nag KM-54 083 strain of sulfur borax (864) is seeded in 1% peptone water at pH 7.6. and grown at 37 ° C for 4 hours. The inoculum is looped to agar pH 7.6. containing infusion of the heart muscle. Grown 18-20 hours at 37 ° C. The biomass is washed off with physiological saline pH - 7.2, washed twice. Rabbits are immunized three times with a three-day interval: the first and second injections are subcutaneously with 20 billion vibrios, and the third - intravenously 5 billion vibrios. On the seventh day the rabbits are exsanguinated. Serum is preserved with sodium thiolate 1: 10,000, diagnostic serum titer is 1: 400, serum is specific, highly active, does not contain heterologous antibodies. 2 tab. B Ј

Description

This invention relates to medical microbiology, namely to the serological identification of pathogenic vibrios.

The aim of the invention is the Vibrio Nag strain of the new 083 serovar used to prepare a monovarial diagnostic serum to identify pathogenic vibrios.

The Vibrio Nag KM-54 (864) 083 strain of serovar is stored in the collection of the Museum of Living Cultures of the All-Union Microbiological Anti-Plague Research Institute under the number CM-54 (864).

The strain Vibrio Nag KM-54 (864) has all the properties characteristic of the genus Vibrio (Table 1).

Morphological signs. Motile, slightly curved sticks, spores and capsules do not form, gram-negative.

Cultural features. On tight A

The agar forms smooth, round transparent colonies with smooth edges with a diameter of 2-2.5 mm. In liquid media causes uniform turbidity, forms a film on the surface.

Physiological properties. Oxidase positive, belongs to the Heiberg group 1, ferments glucose to acid without gas, does not ferment inositol and salicin, forms indole, ferments mannitol, decarboxylates lysine and ornithine, does not hydrogenate arginine, does not phosphorescent, does not grow in 1% peptone water with the addition of 7 and 10% sodium chloride. Not agglutinated with cholera serums 0, Ogawa, Inaba, RO and vibrionic diagnostic serums 02-082 serovars. Not licensed by phages C, Eltor, HDF 3-5,

VI

N) o

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TEPV-2, TEPV-3, TEPV-4, TEPV-5 and TEPV-6. Lysed with phages TEPV-1 and TAPV-7. It is lysed with DDF phages.

It has a specific 0-antigen. When immunizing rabbits with the 0 antigen of this strain, they form specific antibodies.

EXAMPLE 1 To prepare a monovarial diagnostic serum, the 0-antigen of the Vibrio Nag strain KM-54 (864) 083 serovar is used. The culture of the Vibrio Nag KM-54 strain (864) was seeded in 1% peptone water, pH 7.6 and grown at 37 ° C for 4 h, then seeded on freshly mown agar containing infusion of the heart muscle pH 7.6. Crops are incubated at 37 ° C for 18-20 hours. The grown culture is washed from the agar with a physiological solution pH 7.2. The obtained microbial suspension is boiled at 100 ° C for 2 hours from the moment of boiling in a 10% solution of sodium chloride. The suspension is then washed twice with a physiological solution pH 7.2 by centrifugation. After controlling for specific sterility, the microbial suspension is used to immunize rabbits.

Chinchilla rabbits weighing 2-2.5 kg are immunized three times with a three-day interval: the first and second immunizations are made subcutaneously at 20 billion each.

mt, the third injection - 5 billion mt intravenously. On the seventh day after the last injection, the rabbits are exsanguinated. Serum canned by the addition of mer -.

sodium thiolate 1: 10,000. The serum titers are 1: 3200. For diagnostic titer, serum dilution was 1: 400.

Serum is used in the agglutination reaction with vibrios. The agglutination reaction is made using the volumetric method in test tubes. Serum is used in one dilution - diagnostic titer. A physiological solution pH 7.2 was used to dilute the sera and prepare the suspension of the cultures. The reaction with a 2-4 + rating is considered positive.

Serum against vibrios 083 serovar is strictly specific and highly active. A positive reaction was noted only

with homologous strain (table 2).

The use of the proposed serum in the diagnosis of intestinal diseases is of great practical importance, since it expands the possibilities of differentiating pathogenic vibrios.

Claims (1)

Claims of the Invention Bacterial Strain Vibrio Nag 083 KM-54 Serovar (864) Used to Prepare Monovar Diagnostic serum. table 2 The specificity of monovar serum against pathogenic vibrios 083 serovar