RU2421240C1 - Method of producing staphylococcal vaccine - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to microbiology. A vaccine is prepared of 6-8 strains of staphylococci identified as Staphylococcus aureus. The strains are cultivated separately for 14-16 days at temperature 37°C and then at temperature 3-5°C for 6-8 days. Thereafter, the prepared material is mixed and immediately autoclaved at temperature 115-125°C for 15-40 minutes.

EFFECT: vaccine enables intense staphylococci immunity, and also enabling a nonspecific immunity sufficient for treating purulent infection caused by other agents is produced.

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Description

The invention relates to microbiology and can be used to obtain a vaccine intended for the prevention and treatment of purulent infections in animals and humans.

A known method of producing staphylococcal vaccine by prolonged (15-20 days) growing a monovalent staphylococcus culture in broth, followed by filtration or sterilization at 100 ° C (A.M. Bezredka, "Antivirusotherapy", L., 1932).

The disadvantage of this method is that the vaccine made does not take into account the diversity of serological variants of staphylococcus, contains only antigenic components soluble in the medium and, as a result, is not effective enough.

A known method of producing anatoxin vaccine against staphylococcosis of birds, including growing a culture of staphylococci in a nutrient medium, sterilizing biomass followed by detoxification of toxin with formalin, and before growing on medium add 4 vol.% Glycerol, the culture is grown for 7-10 days to a concentration of 5- 7 billion microbial cells in 1 ml, sterilization is carried out at 85-100 ° C for 1-1.5 hours, and detoxification is carried out with 0.6-0.8% formalin solution at an exposure of 45-55 ° C for 7 -10 days (USSR AS No. 1706631, class A61K 39/085 dated 01/17/89, publ. 2 3.01.92, Bull. No. 3).

The disadvantage of this method is that for 7-10 days in the cultivated biomass there is not enough accumulation of staphylococcal metabolism products, and, in addition, formalin treatment gives the vaccine an irritating effect, which is permissible with aerosol spraying of the drug, but undesirable with the intravenous administration of the vaccine to animals.

Closest to the proposed method for producing a staphylococcal vaccine is “A method for producing a staphylococcal and streptococcal polyvalent vaccine” - Vaccinum staphylococcicum (streptococcicum). A suspension of staphylococci or streptococci (1 billion microbial bodies per 1 ml) is killed with formalin (http://www.micro-biology.ru/main-microbiology/medicine/93-vakciny-iz-ubitykh-bakterijj.html). It is used to increase the body's defenses in chronic furunculosis, hydradenitis, sycosis and other staphylococcal and streptococcal skin diseases.

The disadvantage of this method is that the vaccine obtained contains a limited number of antigens - killed microbial cells without biologically active waste products do not contain metabolic inhibitory products of staphylococci (during long-term cultivation in the nutrient medium, specific antimicrobial substances specific only to staphylococcus accumulate), and also contains formalin.

The objective of the invention is to remedy these disadvantages.

The problem is solved as follows.

To obtain a vaccine according to the claimed method take 6-8 strains of staphylococci identified as Staphylococcus aureus. Then, the isolated strains are placed in a standard meat-peptone nutrient broth - each strain in a separate container and cultured for 14-16 days, during which the vaccine acquires antistaphylococcal activity due to the accumulation of metabolic products and antigens of staphylococcus itself, as well as its toxins. Cultivation is carried out at a temperature of 37 ° C.

Then the cultures are kept for 6-8 days at a temperature of 3-5 ° C for the accumulation of more biogenic stimulants in it, similarly to conservation according to the method of V.P. Filatov (BME, third edition, volume 3, p.140), what happens in unfavorable conditions for living cells.

Then the cultures are mixed, poured into vials and immediately autoclaved at a temperature of 115-125 ° C for 15-40 minutes.

The antistaphylococcal activity of the obtained vaccine showed the following results (see table 1).

Table 1 Comparative antistaphylococcal activity of the proposed vaccine and analogue in log Cultivation method Antimicrobial activity Separate cultivation 25.86 Co-cultivation 21.32

The data in table 1 indicate that the antistaphylococcal activity of the obtained vaccine exceeds the activity of the vaccine obtained by co-cultivation.

Treatment of animals with the proposed vaccine is carried out in the same way as treatment with staphylococcal toxoid, but at a dose one and a half times higher, which was established in experiments on immunization of rabbits. The results of the experiments are shown in table 2.

The titer of agglutinins in the blood serum of immunized animals.

table 2 Type of vaccine Agglutinin titer Separate cultivation 1/320 Co-cultivation 1/160

As can be seen from table 2, the proposed vaccine causes a higher titer of growth of antibodies, and therefore has higher immunogenic properties.

Positive example

Received by the proposed method, the vaccine was tested in one of the livestock farms of the Volga region of the Samara region.

The test results are shown in table 3.

Table 3 The results of the therapeutic effect of the vaccine Cow inventory number Diagnosis Treatment result 851 catarrhal mastitis Cured 1437 catarrhal mastitis Cured 1292 serous mastitis Cured

The vaccine was administered to animals subcutaneously, three times, with an interval of one day: on the first day - 3.75 ml, on the second day - 7.5 ml, on the third day - 7.5 ml.

The present invention allows to create a staphylococcal vaccine that provides a sufficiently intense immunity against staphylococci for the treatment of purulent infection, given the fact that the causative agent is the primary immune stimulus. The use of six strains of staphylococci isolated from sick animals in the manufacture of the vaccine allows us to expand the spectrum of action of the vaccine by expanding the range of antigenic determinants of both staphylococcus and its vital products. The use of autoclaving for sterilization of the vaccine at the indicated parameters allows, in addition to sterilization, cell destruction and protein denaturation, as a result of which the vaccine also acquires non-specific properties and can be used to treat purulent infections caused by other pathogens. Due to the fact that the vaccine has a nonspecific effect on the body, the determination of the etiotropic pathogen is not mandatory. Since the proposed vaccine does not contain formalin, it can be administered both intramuscularly and subcutaneously. Separate cultivation of staphylococcus strains increases its antimicrobial, immunogenic properties, therapeutic effect and is more economical.

Claims (1)

A method of obtaining a staphylococcal vaccine for the treatment and prevention of purulent infections, comprising growing six to eight strains of pyogenic staphylococci in a liquid nutrient medium and sterilizing biomass, characterized in that the cultivation of six to eight strains of pyogenic staphylococcus identified as Staphylococcus aureus is carried out separately for 14 16 days at a temperature of 37 ° C, then cultivated separately at a temperature of 3-5 ° C for 6-8 days, after which the resulting material is mixed and immediately autoclaved at a temperature of 115- 125 ° C for 15-40 minutes