CN101463374A - Bacillus dysenteriae diagnose reagent - Google Patents

Abstract

The invention relates to a dysentery bacillus diagnostic reagent. The main components comprise tryptone, sodium citrate, sodium deoxycholate, monopotassium phosphate, dipotassium hydrogen phosphate, sodium chloride, calf serum, glucose and mannite which are processed to obtain the reagent by certain technology. The dysentery bacillus diagnostic reagent is used for feces enriched culture dysentery bacillus. The key to identifying the sort of bacteria by culture experiment is accuracy, high-speed and simpleness and convenience. The bacterination volume can affect the detection result directly. The enriched culture is always required to be carried out on a sample by different methods to identify the sort of the bacteria, which is the measure used most frequently in a microbiological laboratory. Due to the limit of the application range of the current culture condition, diagnostic reagent used for feces dysentery bacillus enriched culture is not specific and has the problems of large required inoculation volume, long culture time and proneness of showing false positive or false negative, etc. The dysentery bacillus diagnostic reagent has the advantages as follows: (1) the dysentery bacillus diagnostic reagent requires small inoculation volume and short culture time and can breed fast just by a small amount of bacterium; the result can be observed just after the culture for 5 to 6 hours at the normal room temperature or 37 DEG C; (2) the false positive or the false negative does not occur easily; and (3) the dysentery bacillus diagnostic reagent can destroy the bacteriostatic action of acheomycin, aureomycin, terramycin, neomycin, polymyxin and streptomycin existing in the sample.

Description

Bacillus dysenteriae diagnose reagent

Technical field the present invention relates to a kind of bacillus dysenteriae diagnose reagent, and its main component is that Tryptones, Sodium Citrate, sodium deoxycholate, potassium primary phosphate, dipotassium hydrogen phosphate, sodium-chlor, calf serum, glucose, N.F,USP MANNITOL are made through certain technology.Use for ight soil and increase bacterium cultivation dysentery bacterium.Utilize culture experiment to differentiate the kind of bacterium, it is accurate, quick, easy that its key is.And the bacterial load size can directly influence detected result.Often need that sample is increased bacterium by different methods and cultivate, carry out the discriminating of bacterial species, this is the most frequently used means of present Microbiological Lab.Because at present the range of application of culture condition is limit, increasing the diagnostic reagent that bacterium cultivates for the ight soil dysentery bacterium does not have specific, has that required inoculum size is big, incubation time length, is prone to problems such as false positive or false negative.Along with the progress of biological study, increase the research of bacterium diagnostic reagent also in constantly developing to what bacterium was differentiated both at home and abroad.Biological worker wishes to have always and a kind ofly can overcome above-mentioned insufficient bacterium the diagnostic reagent----bacillus dysenteriae diagnose reagent that increases.

Background technology the object of the present invention is to provide and a kind ofly can overcome the existing insufficient bacillus dysenteriae diagnose reagent of differential diagnosis reagent.Similar diagnostic reagent with other is compared possesses following characteristics: required inoculum size is little, and a small amount of bacterium can breed rapidly, under room temperature usually or 37 ℃ cultivate and got final product observations in 5-6 hour; Be not prone to false positive and false negative; Can destroy the antibacterial substance that exists in the sample, bacterial growth is not suppressed.

Summary of the invention main points of the present invention are to select suitable component, and rationally are mixed, through heat, dissolve, technology such as mixing, cooling, packing, high-temperature sterilization forms a kind of bacillus dysenteriae diagnose reagent.

The prescription following (component and weight percent % thereof) that the present invention selects: Tryptones (1.8-2.2), Sodium Citrate (0.4-0.5), sodium deoxycholate (0.04-0.06), potassium primary phosphate (0.14-0.16), dipotassium hydrogen phosphate (0.38-0.42), sodium-chlor (0.4-0.5), glucose (0.10-0.12), N.F,USP MANNITOL (0.15-0.25), calf serum (0.4-0.5), distilled water add to 100ml.

This diagnostic reagent contains Sodium Citrate and sodium deoxycholate, and Gram-negative bacteria is had restraining effect, intestinal bacteria, and Pseudomonas aeruginosa and Bacillus proteus, poor growth in inoculating 6 hours, and dysentery bacterium can relatively obtain propagation. sodium-chlor is used to adjust osmotic pressure; Potassium primary phosphate is used to regulate pH.Tryptones, calf serum, glucose, N.F,USP MANNITOL are the nutrition in the diagnostic reagent and support composition.Be computing time after the specimen inoculation, under common room temperature or 37 ℃ of cultivations, the bacterium of increasing effect arranged all. increase bacterium and cultivate after 6 hours, promptly with the method for scoring culture transferring to the enteron aisle selectivity diagnostic reagent to separate pathogenic bacterium.

The preparation method of product of the present invention is:

1), Tryptones, Sodium Citrate, sodium deoxycholate, potassium primary phosphate, dipotassium hydrogen phosphate, sodium-chlor, glucose, N.F,USP MANNITOL are added in the distilled water in proportion, the reheat dissolving is corrected pH to 7.0 and is corrected pH to 6.8, uses filter paper filtering;

2), pressuresteam sterilization 0.70kg15 minute;

3), under the aseptic condition, add aseptic calf serum, packing ampere bottle, every bottle of about 5ml is stored in the refrigerator standby.

The present invention has the following advantages: (1) required inoculum size is little, and incubation time is short, and a small amount of bacterium can breed rapidly, under room temperature usually or 37 ℃ cultivate and got final product observations in 5-6 hour; (2) be not prone to false positive or false negative; (3) can destroy the bacteriostatic action of the tsiklomitsin, duomycin, terramycin, Xin Meisu, polymyxin and the Streptomycin sulphate that exist in the sample.

The present invention is described in further detail below in conjunction with embodiment for embodiment:

Be configured according to following table institute column data (weight percent %) and described step thereof:

Prescription one:

Tryptones 1.8-2.2

Sodium Citrate 0.4-0.5

Sodium deoxycholate 0.04-0.06

Potassium primary phosphate 0.14-0.16

Dipotassium hydrogen phosphate 0.38-0.42

Sodium-chlor 0.4-0.5

Glucose 0.10-0.12

N.F,USP MANNITOL 0.15-0.25

Calf serum 0.4-0.5

Distilled water adds to 100

Prescription two:

Tryptones 1.8-2.0

Sodium Citrate 0.4-0.45

Sodium deoxycholate 0.04-0.05

Potassium primary phosphate 0.14-0.15

Dipotassium hydrogen phosphate 0.38-0.40

Sodium-chlor 0.4-0.45

Glucose 0.10-0.11

N.F,USP MANNITOL 0.15-0.20

Calf serum 0.4-0.45

Distilled water adds to 100

Prescription three:

Tryptones 2.0-2.2

Sodium Citrate 0.45-0.5

Sodium deoxycholate 0.05-0.06

Potassium primary phosphate 0.15-0.16

Dipotassium hydrogen phosphate 0.40-0.42

Sodium-chlor 0.45-0.5

Glucose 0.11-0.12

N.F,USP MANNITOL 0.18-0.25

Calf serum 0.45-0.5

Distilled water adds to 100

Prescription four:

Tryptones 1.9-2.1

Sodium Citrate 0.42-0.45

Sodium deoxycholate 0.05-0.06

Potassium primary phosphate 0.15-0.16

Dipotassium hydrogen phosphate 0.39-0.40

Sodium-chlor 0.42-0.45

Glucose 0.11-0.12

N.F,USP MANNITOL 0.17-0.22

Calf serum 0.42-0.45

Distilled water adds to 100.

Claims (2)

1, a kind of bacillus dysenteriae diagnose reagent is to be main component with Tryptones, Sodium Citrate, sodium deoxycholate, potassium primary phosphate, dipotassium hydrogen phosphate, sodium-chlor, calf serum, glucose, N.F,USP MANNITOL; It is characterized in that having following prescription (component and weight percent % thereof): Tryptones (1.8-2.2), Sodium Citrate (0.4-0.5), sodium deoxycholate (0.04-0.06), potassium primary phosphate (0.14-0.16), dipotassium hydrogen phosphate (0.38-0.42), sodium-chlor (0.4-0.5), glucose (0.10-0.12), N.F,USP MANNITOL (0.15-0.25), calf serum (0.4-0.5), distilled water 100ml.

2, the preparation method of the described bacillus dysenteriae diagnose reagent of a kind of claim 1 is characterized in that having following step:

1), Tryptones, Sodium Citrate, sodium deoxycholate, potassium primary phosphate, dipotassium hydrogen phosphate, sodium-chlor, glucose, N.F,USP MANNITOL are added in the distilled water in proportion, the reheat dissolving is corrected pH to 7.0 and is corrected pH to 6.8, uses filter paper filtering;

2), pressuresteam sterilization 0.70kg15 minute;

3), under the aseptic condition, add aseptic calf serum, packing ampere bottle, every bottle of about 5ml is stored in the refrigerator standby.