Indicator paper systems for identification of microorganisms. Set No. 2 for Intergenerational
and species differentiation of enterobacteria
Form of issue
Issue in a set of 13 tests: 12 vials with NIB-disks, 2 tubes with NIB-strips.
Composition
Indicator paper systems (NIBs) for identification of microorganisms (diagnostic kit) are discs or strips of chromatographic paper containing certain amounts of substrate in combination with an indicator stabilized by a film-forming coating - polyvinyl alcohol.
NIB is released in a set of 13 tests: 12 vials with SIB disks (NIB with sorbitol, inositol, lysine, ornithine, sodium citrate, sodium malonate, for determination of galactosidase, urease, phenylalanine deaminase (1 vial of NIB with phenylalanine, 1 vial of NIB with ferric chloride), hydrogen sulphide, for the Foges-Prosvauer reaction) and 2 tubes with NIB strips (NIB for the determination of oxidase and indole). NIB-disks are produced in vials, NIB-strips in test tubes in a cardboard bundle with instructions for use.
Set No. 2 provides 50 tests.
Indications for use
Determination of the enzymatic activity of microorganisms of the Enterobacteriaceae family and their identification.
Dosage regimen and method of administration
EQUIPMENT AND REAGENTS
• Thermostat providing a temperature (37 ± 1) ° С;
• Autoclave;
• Tubes of glass;
• Petri dishes;
• Tweezers;
• Bacteriological loop;
• Glass wand;
• Medical cotton wool hygroscopic;
• Sodium chloride solution 0.9% pH 7.3 ± 0.1 sterile;
• Phosphate-buffered saline solution pH 5.5 ± 0.2 (TU 9389-117-14237183-08);
• Vaseline sterile oil;
• α-naphthol alcohol solution 6%;
• Sodium hydroxide solution is 40%.
ANALYZED SAMPLES
Objects of research in sanitary and clinical microbiology.
CONDUCTING ANALYSIS
The main requirements for working with a set of reagents
"Indicator paper systems for identification of microorganisms. Set No. 2 for intergeneric and species differentiation of enterobacteria »:
- Studies are conducted with a clean culture, as well as with individual colonies
directly from cups with differential diagnostic environments (Endomo, Ploskireva, bismuth-sulfite agar medium);
- immersion of discs in test tubes is made with burnt tweezers;
- to obtain clear results, compliance with the temperature
thermostat mode (37 ± 1) ° С, the pH of the nutrient media used and the treatment mode of the dishes.
Determination of oxidase activity.
The culture grown on the surface of the nutrient agar for culturing microorganisms for 18-24 hours at a temperature of (37 ± 1) ° C is triturated with a platinum loop or glass rod on a test strip of NIB placed in a Petri dish. On one strip you can study 10-14 cultures (no more).
Determination of the utilization of carbohydrates and polyhydric alcohols.
In a test tube with 0.3 ml of sterile sodium chloride solution of 0.9% pH 7.3 ± 0.1, a complete loop of culture grown on the surface of nutrient agar for culturing microorganisms is suspended for 18-24 hours at a temperature of (37 ± 1) ° FROM. Then immerse the NIB-disk with the corresponding carbohydrate or polyhydric alcohol. The medium in the test tubes becomes red as a result of the rapid diffusion of the indicator into it. If it is necessary to take into account gas formation, it is recommended to use a small lump of sterile absorbent cotton, which is placed in a test tube. As a control, NIB disks immersed in test tubes with sterile sodium chloride solution of 0.9% pH 7.3 ± 01 serve as a control. The tubes are incubated at a temperature of (37 ± 1) ° C.
Determination of activity - galactosidase.
In a test tube with 0.3 ml of sterile sodium chloride solution of 0.9% pH 7.3 ± 0.1, a complete loop of culture grown on the surface of nutrient agar for culturing microorganisms is suspended for 18-24 hours at a temperature of (37 ± 1) ° C, then the NIB-disk is placed in this suspension. As a control, a NIB disk is used, placed in a test tube with sterile sodium chloride solution of 0.9%. Incubate at a temperature of (37 ± 1) ° C.
Definition of indole formation.
In a test tube with 0.3 ml of sterile sodium chloride solution of 0.9% pH 7.3 ± 0.1, a complete loop of culture grown on the surface of nutrient agar for culturing microorganisms is suspended for 18-24 hours at a temperature of (37 ± 1) ° FROM. The strip for determining the indole is folded along the line planned for it by a factor of two and pincers are lowered to the bottom of the tube so that the long colorless end is immersed in the suspension of the culture and the short colored end is above the surface of the suspension. As a control, use a strip placed in a test tube with sterile sodium chloride solution of 0.9% pH 7.3 ± 0.1. Incubate at a temperature of (37 ± 1) ° C.
Determination of urease activity.
In a test tube with 0.3 ml of sterile sodium chloride solution of 0.9% pH 7.3 ± 0.1, a complete loop of culture grown on the surface of nutrient agar for culturing microorganisms is suspended for 18-24 hours at a temperature of (37 ± 1) ° C, then a SIB disk with urea is dipped into this slurry.