CN101463375A - Improved bacteria differential diagnosis agent - Google Patents

Abstract

The invention relates to a modified bacteria identification diagnostic reagent. The main components comprise sodium citrate, sodium chloride, magnesium sulfate, monopotassium phosphate, monoammonium phosphate, thymolsulfonphthalein, calf serum and agar which are processed to obtain the reagent by certain technology. The modified bacteria identification diagnostic reagent is used for identifying whether citrate can be applied to bacteria and is used for identifying the sort of the bacteria by a biochemical reaction experiment exactly, fast and simply. Different bacterium have a respective unique enzyme system, so that decomposed and synthesized metabolites produced during the metabolizing process are different; the decomposed and synthesized metabolites have different biochemical characteristics; and the biochemical reaction which is generated under corresponding culture condition by the different biochemical characteristics is used for identifying the sort of the bacteria, which is the measure used most frequently in a microbiological laboratory. The modified bacteria identification diagnostic reagent has the advantages as follows: (1) the modified bacteria identification diagnostic reagent requires small inoculation volume, produced alkali can discolor just by few bacterium; (2) an indicator discolors fast and obviously and the result can be observed within 24 hours; (3) the modified bacteria identification diagnostic reagent does not show false positive easily and only allows the bacterium which can use the citrate to grow and discolor; (4) the modified bacteria identification diagnostic reagent does not show false negative easily and the magnesium sulfate can destroy the bacteriostatic action of acheomycin, aureomycin, terramycin, neomycin, polymyxin and streptomycin existing in a sample; and (5) experiment positive control bacteria increases and breeds fast in a solid culture medium, and the produced alkali leads the culture medium to discolor.

Description

The improvement bacteria differential diagnosis agent

Technical field the present invention relates to a kind of improvement bacteria differential diagnosis agent, and its main component is Sodium Citrate, sodium-chlor, sal epsom, potassium primary phosphate, primary ammonium phosphate, bromothymol blue, calf serum, agar, makes through certain technology.In order to differentiate that can bacterium utilize citrate.Utilize the biochemical reaction test to differentiate the kind of bacterium, it is accurate, quick, easy that its key is.Because different bacterium has its unique enzyme system separately, thereby the decomposition that is produced in metabolic process is also different with the synthetic meta-bolites, these decomposition or synthetic meta-bolites respectively have different biochemical characteristics again, the biochemical reaction that utilizes its different biochemical characteristics under corresponding culture condition, to be produced again, 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 and indicator is limit, the observation bacterium exists to the discriminating differential diagnosis agent of organic acid salt and ammonium salt utilization that required inoculum size is big, incubation time is long, indicator discoloration is not obvious, be prone to problems such as false positive or false negative in the biochemical reaction test.Can the improvement bacteria differential diagnosis agent be to observe the comprehensive differential diagnosis agent of bacterium to organic acid salt and ammonium salt utilization, utilize citrate in order to the discriminating bacterium, thereby further judge kind and the biochemical characteristics of bacterium.Along with the progress of biological study, the domestic and international research that bacterium is differentiated is also in constantly developing.Biological worker wishes to have always and a kind ofly can overcome above-mentioned insufficient differential diagnosis agent----improvement bacteria differential diagnosis agent.

Background technology the object of the present invention is to provide and a kind ofly can overcome the existing insufficient improvement bacteria differential diagnosis agent of differential diagnosis agent.Similar differential diagnosis agent with other is compared possesses following characteristics: required inoculum size is little, and the alkali variable color can appear producing in a small amount of bacterium; Indicator discoloration is obvious fast, gets final product observations in 24 hours; Be not prone to false positive, only for the bacterium that can utilize Sodium Citrate can grow, variable color; Be not prone to false negative, can destroy the antibacterial substance that exists in the sample, bacterial growth is not suppressed; Test positive control bacterium increases the bacterium breeding rapidly in this solid differential diagnosis agent, and product alkali makes differential diagnosis agent variable color.

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 improvement bacteria differential diagnosis agent.

The prescription following (component and weight percent % thereof) that the present invention selects: Sodium Citrate (0.3-0.7), sodium-chlor (0.3-0.7), sal epsom (0.01-0.03), potassium primary phosphate (0.08-0.12), primary ammonium phosphate (0.08-0.12), bromothymol blue (0.8-1.2), calf serum (2.5-3.0), agar (1.5-2.5), distilled water add to 100.

Sodium Citrate is unique source of carbon in the differential diagnosis agent, and primary ammonium phosphate is unique source of nitrogen in the differential diagnosis agent.Can utilize Sodium Citrate can grow in the differential diagnosis agent for the bacterium of carbon source, and decompose citrate and produce carbonate at last, make the differential diagnosis agent become alkalescence, the bromothymol blue indicator in differential diagnosis agent this moment becomes mazarine by green.Sal epsom can destroy the bacteriostatic action of the tsiklomitsin, duomycin, terramycin, Xin Meisu, polymyxin and the Streptomycin sulphate that exist in the sample; Sodium-chlor is used to adjust osmotic pressure; Potassium primary phosphate is used to regulate pH.Calf serum, agar are the nutrition in the differential diagnosis agent and support composition; Test positive control bacterium increases the bacterium breeding in this solid differential diagnosis agent, and product alkali makes differential diagnosis agent variable color.

The preparation method of product of the present invention is:

1), Sodium Citrate, sodium-chlor, sal epsom, potassium primary phosphate, primary ammonium phosphate, agar are added in the distilled water in proportion, pH to 6.8 is corrected in the reheat dissolving, filters with flannelette;

2) add the bromothymol blue mixing, again, pressuresteam sterilization 1.05kg15 minute;

3), under the aseptic condition, add aseptic calf serum, the packing test tube, the about 2ml of every pipe will contain the sterilization test tube inclination certain angle of differential diagnosis agent while hot, and bevel after the condensation is stand-by.

The present invention has the following advantages: (1) required inoculum size is little, and the alkali variable color can appear producing in a small amount of bacterium; (2) indicator discoloration is obvious fast, gets final product observations in 24 hours; (3) be not prone to false positive, only for the bacterium that can utilize Sodium Citrate can grow, variable color; (4) be not prone to false negative, sal epsom can destroy the bacteriostatic action of the tsiklomitsin, duomycin, terramycin, Xin Meisu, polymyxin and the Streptomycin sulphate that exist in the sample; (5) test positive control bacterium increases the bacterium breeding rapidly in this solid differential diagnosis agent, and product alkali makes differential diagnosis agent variable color.

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:

Sodium Citrate 0.3-0.7

Sodium-chlor 0.3-0.7

Sal epsom 0.01-0.03

1% bromothymol blue spirituous solution 0.8-1.2

Potassium primary phosphate 0.08-0.12

Primary ammonium phosphate 0.08-0.12

Calf serum 2.5-3.0

Agar 1.5-2.5

Distilled water adds to 100

Prescription two:

Sodium Citrate 0.3-0.6

Sodium-chlor 0.3-0.6

Sal epsom 0.01-0.02

1% bromothymol blue spirituous solution 0.8-1.1

Potassium primary phosphate 0.08-0.11

Primary ammonium phosphate 0.08-0.11

Agar 1.5-2.2

Calf serum 2.5-2.8

Distilled water adds to 100

Prescription three:

-Sodium Citrate 0.4-0.7

Sodium-chlor 0.4-0.7

Sal epsom 0.02-0.03

1% bromothymol blue spirituous solution 0.9-1.2

Potassium primary phosphate 0.09-0.12

Primary ammonium phosphate 0.09-0.12

Calf serum 2.7-3.0

Agar 1.8-2.5

Distilled water adds to 100

Prescription four:

Sodium Citrate 0.4-0.6

Sodium-chlor 0.4-0.6

Sal epsom 0.01-0.03

1% bromothymol blue spirituous solution 0.9-1.1

Potassium primary phosphate 0.09-0.11

Primary ammonium phosphate 0.09-0.11

Agar 1.8-2.2

Calf serum 2.6-2.8

Distilled water adds to 100.

Claims (2)

1, a kind of improvement bacteria differential diagnosis agent, its main component is Sodium Citrate, sodium-chlor, sal epsom, potassium primary phosphate, primary ammonium phosphate, bromothymol blue, calf serum, agar, makes through certain technology; It is characterized in that having following prescription (component and weight percent % thereof): Sodium Citrate (0.3-0.7), sodium-chlor (0.3-0.7), sal epsom (0.01-0.03), potassium primary phosphate (0.08-0.12), primary ammonium phosphate (0.08-0.12), bromothymol blue (0.8-1.2), calf serum (2.5-3.0), agar (1.5-2.5), distilled water add to 100.

2, the preparation method of the described improvement bacteria differential diagnosis agent of a kind of claim 1 is characterized in that having following step:

1), Sodium Citrate, sodium-chlor, sal epsom, potassium primary phosphate, primary ammonium phosphate, agar are added in the distilled water in proportion, pH to 6.8 is corrected in the reheat dissolving, filters with flannelette;

2) add the bromothymol blue mixing, again, pressuresteam sterilization 1.05kg15 minute;

3), under the aseptic condition, add aseptic calf serum, the packing test tube, the about 2ml of every pipe will contain the sterilization test tube inclination certain angle of substratum while hot, and bevel after the condensation is stand-by.