RU187404U1 - DEVICE FOR COLLECTION OF POINT AND SMALL COLONIES OF MICRO-ORGANISMS FROM THE SURFACE OF DENSE NUTRIENTS - Google Patents

Abstract

The utility model relates to the field of medicine and biotechnology and can be used to select colonies of microorganisms from solid nutrient media for the collection of necessary biomass and further identification using mass spectrometry. The proposed version of the device for collecting point and small colonies of microorganisms from the surface of solid nutrient media is a circular platform, on the front of the platform there is a handle, at the end of which there is an octagonal section device, for more convenient grip with fingers; on the inside of the platform there are two boots, intersecting in the center of the platform at an angle of 90 °, the corner at the base of the “boot”, creating a rise of 9 ° 25 ', on the back of the boot there is a rounding, the “boot” is attached to the platform using the “legs” , the rib of the outer part of the platform has a rounding, all parts of the device are made of optical polystyrene or general polystyrene, in the case of manufacturing on a 3D printer, ABS, PLA, HIPS are allowed. The utility model allows you to collect the necessary biomass for further identification using the MALDI-TOF mass spectrometry method. 1 ill.

Description

The utility model relates to the field of medicine and biotechnology and can be used to select colonies of microorganisms from solid nutrient media for the collection of necessary biomass and further identification using the MALDI-TOF mass spectrometry method.

In connection with the rapid pace of development of modern microbiology and the emergence of new methods for the exact species identification of microorganisms, new methods of sample preparation and selection of colonies of microorganisms for subsequent identification are also required. One of the most significant and promising methods from the point of view of quick and high-quality identification is the MALDI-ToF mass spectrometry method. In this case, the research material is a pure culture of bacteria collected directly from solid nutrient media. In this case, a pure culture can be represented as a single colony, as well as a bacterial mass, if the colonies are small in size.

Identification of most microorganisms is possible without preliminary preparation of material for research, however, there is a fairly wide group of microorganisms for which sample preparation procedures are necessary, due to the peculiarities of their structure. These include microorganisms such as slowly growing non-tuberculous mycobacteria, streptomycetes, actinomycetes and other members of the Actinomycetales family. The microorganisms described above during cultivation on solid nutrient media often give rise in the form of point and small colonies with a diameter of up to 2 mm; therefore, their identification is rather difficult due to insufficient biomass of bacteria necessary for identification. Also, in the process of sample preparation of biomaterial, it is necessary to minimize the ingress of components of nutrient media into the microbial mass, since they reduce the quality of identification.

Standard devices for collecting microbial cultures from culture media are bacteriological loops of various diameters and spatulas of various sizes. The disadvantage of these devices is that they do not allow to collect the biomass volume of microorganisms necessary for identification, giving small point colonies and can capture fragments of dense nutrient media.

The purpose of creating a useful model is to develop a device for collecting point and small colonies of microorganisms from the surface of dense nutrient media, which will allow you to collect the necessary biomass for further identification using the MALDI-ToF mass spectrometry method.

Brief description of the drawing. The drawings attached to the description provide a design diagram of a device for collecting point and small colonies of microorganisms from the surface of a dense nutrient medium. Figure 1 - a device for collecting point and small colonies of microorganisms from the surface of dense nutrient media in two projections: 1) front view with the direction of view (A and B with arrows) for the image of the projection of the top view and bottom view (projection A and B) , 2) a front view of a fragment of a platform with a handle in a 2: 1 scale (projection B).

In the drawing, Figure No. 1. The following items are indicated: 1 - platform of circular cross-section, 2 - adaptation of an octagonal cross-section on the handle.

Implementation of a utility model. The proposed version of the device for collecting point and small colonies of microorganisms from the surface of solid nutrient media, is a circular platform (position 1 in Fig. 1) with a diameter of 5.00 mm to 5.20 mm and a height of 2.0 mm to 2, 1 mm. On the front of the platform there is a handle with a height of 59.5 mm to 60 mm, at the end of which there is an octagonal section fixture (position 2 in Fig. 1) for finger grip. The height of the device (position 2 in Fig. 1) is from 14.8 mm to 15.2 mm, the distance from the center to the wide edge is from 0.9 mm to 1.0 mm, the distance from the center to the narrow edge is from 0.8 mm to 0.9 mm. The width of the wide face is from 0.9 mm to 1.0 mm; the width of the narrow face is from 0.5 mm to 0.6 mm. On the inside of the platform are two boots intersecting in the center of the platform at an angle of 90 °. The base in the form of a “boot” has a length of 2.4 mm to 2.6 mm, the angle at the base of the “boot” creating a lift is 9 ° 25 '. On the back of the boot there is a rounding of R0.5 mm. The “boot” is attached to the platform with the help of a “leg” with a height of 0.6-0.05 mm. The rib of the outer part of the platform has a rounding of the outer diameter of R0.5 mm.

The device is made of optical polystyrene or general purpose polystyrene GOST 20282-86.

The utility model allows you to collect the necessary biomass of point and small colonies of microorganisms from the surface of solid nutrient media for further identification using the MALDI-ToF mass spectrometry method.

The technical result of the claimed utility model, which consists in ensuring the collection of the necessary biomass for further identification using the MALDI-ToF mass spectrometry method, is confirmed by the data obtained from testing a utility model prototype with colony collection of 30 strains of known microorganisms and their subsequent identification.

The experiment included 30 strains of slowly growing non-tuberculous mycobacteria. Using a bacteriological loop and a prototype, it was possible to collect material from culture media for all tested strains.

The selected material was subjected to an extraction process followed by the identification of microorganisms. Identification was carried out using MALDI-ToF mass spectrometry, according to which species identification is evaluated by Score: more than 2 (species identification), Score from 1.7 to 2 (identification to genus), Score lower; 1.7 (microorganism not identified).

As a result of the study of strains selected using the bacteriological loop, 33.3% were identified with a Score greater than 2, 66.7% with a Score lower than 1.7 - no identification was performed. As a result of the identification of strains selected using a prototype of the claimed utility model, 80.0% of strains with a Score greater than 2, 20% with a Score from 1.7 to 2 were identified.

The studies described above confirm the technical result of the claimed utility model, which consists in ensuring the collection of the necessary biomass for further identification using the MALDI-ToF mass spectrometry method.

INFORMATION SOURCES:

1. Clinical laboratory diagnostics: national leadership: in 2 volumes - T.P. / Ed. V.V. Dolgova, V.V. Menshikov. - M.: GEOTAR-Media, 2013 .-- 808 p.

2. Guide to medical microbiology. General and sanitary microbiology. Book I / Coll. authors // Ed. A.S. Labinskaya, E.G. Volina. - M.: Publishing house BINOM, 2008. - 1080 p.

3. Methods of general bacteriology: Per. from English / Ed. F. Gerhardt et al. - Moscow: Mir, 1983 .-- 536 p.

Claims (1)

A device for collecting point and small colonies of microorganisms from the surface of a dense nutrient medium, characterized in that it is a circular platform, where on the front of the platform there is a handle, at the end of which there is an octagonal section for gripping with fingers, and on the inside of the platform two “boots” with a length of 2.4 to 2.6 mm, intersecting in the center of the platform at an angle of 90 °, and the angle at the base of the “boot” creating a lift is 9 ° 25 ', in addition, on the back apozhka has a rounding, "boot" is secured to the platform by means of the "legs", the outer edge of the platform has a rounding, and all parts of the device are made of polystyrene.

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