SU1028718A1 - Strain 97 of phage nag-vibrios 0-41 - Google Patents

Abstract

Strain 97

Description

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This invention relates to medical microbiology and can be used in laboratory diagnostics. . A known strain of Phagum NAG-vibrionom O-lnefotype 88, active in the removal of NAG-vibrios of serotype. However, phage 83 is characterized by low stability during storage even at A-10 C, In these conditions, the concentration of living particles to 2 may. storage decreases with 10 times. At a higher temperature (37 °), the phage dies after 5 days. Thus, phage 88 preparations have a shelf life of no more than two months. The low viability of phage 88 limits its scope "- (and for diagnostic purposes. The purpose of the invention is a new strain of specific phage - to identify enteropathogenic NAG-vibrios of serovar O-I, suitable for widespread use in medicine,. The proposed strain was isolated from water open reservoirs on the territory of the USSR As a result, a moderate phage 1203 was obtained from the culture of NAG-ribrion 1. serovar, from which a virulent mutant was isolated on a lawn indicator strain, 2 0-41 Serova 1p. Strain 97 (1203-2) phage NAG-vibrio serov The Phagura MAG 97 ovara, which is specifically active against NAG-vibrios 0-41 serovars, is stored in the collection of the Tbilisi Research Institute of Vaccines and Serums No. F-100. The resulting strain 97 of the NAG vibrios 0-41 serovar phage has the following morphological and physiological characteristics. of colonies on the lawn of NAG-vibrio 442 grown on Martin's agar is equal to 3.4 tO, 4 mm in diameter. The edge of the colonies is even, the bottom is transparent, and along the periphery there is a zone of incomplete lysis. Unlike phage 88, attributed to the X serotype of NAG-vibrios 0-41 serovar phages, phage 97 is related to the previously unknown serotype of the VAG-phage, which is designated as TT eurotype phage of NAG-vibrioes 0-41 serovar with the known 10 serotypes of cholera phages and NAG-phage 88 has no serological relationship. Its corpuscles have a hexagonal head with the size of 5В21Я and a long elastic tail process without a cover. The process length is 171 D. When laboratory testing the properties of phage 97, it is established that it, like phage 88, has a wide range of specific effects on serovar vibrios, but differs from phage 88 in serological properties, particle structure and stability during storage. Phage 97 preparations, at a concentration of particles VI-10, lysed 81 strains of O-il saltwater, not more than 2.2% of NAG-vibrios of the other 5 serovars, keeping their titer f (concentration of living particles) unchanged for 2 years of storage at 4-10 ° C and at least 2 wt. at 37 ° C, non-choleric vibrios (phosphorescent), microorganisms of the genus Pseudomonas Acromonas and the intestinal group are not sensitive to the action of phage 97. Phage 97 is actively reproduced on NAG-vibrio 2 cells in Martin's broth, pH 7.6. At a seeding dose of 1 ° microbial cells per 1 ml of medium and a multiplicity of infection of 1: 1 during incubation for 2 h 30 min at 37 ° C, the phage accumulates up to 10% of particles per ml of medium. To obtain a diagnostic preparation of phage 97, agar and Martin broth with a pH of 7.6 are used. The culture of the producer strain No. 442, an 18-hour growth at 37 on Martin's agar, is seeded in Martin's broth. After 3 hours of growth at 37 ° C, the broth culture is added to the nutrient medium (Martin broth) at a rate of 110 mt. per ml, phage 97 are simultaneously introduced at the rate of infection multiplicity of 1: 1. After 2 hours and 30 minutes of cultivation, the phagolysate is filtered through bacterial -. ny ltry brand f-3, f-5. The filtrate is poured into ampoules and monitored for specific and general sterility and activity. The finished product is used for the diagnosis of the following method. Prepare a lawn of the test strain in a soft Gracia agar layer, i.e. O, 2 ml of broth culture of the test strain is added to 5 ml of O, molten agar (45 ° C), mixed and poured onto an agar (2%) plate in a Petri dish. After the second layer hardens, phage 97 is dripped onto its surface. placed in a thermostat at. Recording of results is carried out through H-8 hours. Finding at the site of application of the phytic spot, the strain under study is attributed to the 0-4 l serovar vibrios.

 Phage 97 is highly stable when stored not only under the conditions of the refrigerator, but also in adverse temperature conditions {374).

(In the f-IOC refrigerator) its titer remains unchanged for at least 2 years (observation period), only after 3 months. storage, a decrease in the concentration of particles was noted by a factor of 10, whereas phage 88 at

/ Dies after 5 days, and if the diagnostic titer is maintained for not more than 2 months. Unlike phage 88, which can be used for diagnosis in a very limited number of institutions that have the conditions and resolution for its reproduction every 2 months.

phage. 97 can be prepared l Specialized bath, in advance in one of the specialized laboratories and | Assigned for practical use in peripheral health facilities.

The use of phage 97 will lead to a cessation of the flow of nutrient media and working time for the preparation of a diagnostic preparation used to identify NAG vibrios of the serovar. A wider application of the diagnostic method based on the use of phage 97- will improve the diagnosis of vibrios.

The use of phage 97 in laboratory practice will eliminate the need for setting serological reactions, which are currently the only way to identify NAG-vibrios of certain serotypes.

Claims (1)

Strain 97 (1203/442) of the NAG vibrio phage 0-41 serovar Rkadit ΝΑΟ-97> specifically active about the NAG vibrio ratio 0-41 serovar, is stored in the collection of the Tbilisi Research Institute of Vaccines and Serums under the number f-100. ι 1028718 · 2