CN106701892A - TTC (2,3,5-triphenyltetrazolium chloride) chromogenic culture medium for drug sensitivity tests, drug sensitivity kit, preparation method of drug sensitivity kit and drug sensitivity test method - Google Patents
TTC (2,3,5-triphenyltetrazolium chloride) chromogenic culture medium for drug sensitivity tests, drug sensitivity kit, preparation method of drug sensitivity kit and drug sensitivity test method Download PDFInfo
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Abstract
The invention provides a TTC (2,3,5-triphenyltetrazolium chloride) chromogenic culture medium for drug sensitivity tests. The TTC chromogenic culture medium comprises BHI (brain heart infusion) broth, acid-hydrolyzed protein, soluble starch and TTC. The invention further provides a drug sensitivity kit. The drug sensitivity kit comprises a kit part and a drug-sensitive plate part, wherein the kit part comprises the TTC chromogenic culture medium. The invention further provides a preparation method of the drug sensitivity kit. The preparation method comprises preparation of the drug-sensitive plate part and preparation of the TTC chromogenic culture medium, and the drug-sensitive plate part is provided by the invention. The invention further provides a drug sensitivity test method. The drug sensitivity test method comprises steps of diluting a bacterial suspension with the culture medium, adding the diluted bacterial suspension to drug-sensitive holes and performing culture; the culture medium is the TTC chromogenic culture medium. With the adoption of the TTC chromogenic culture medium, the drug-sensitivity kit, the drug sensitivity test method and the drug sensitivity kit prepared with the preparation method of the drug sensitivity kit, drug sensitive test results can be directly and easily observed, deviation cannot be caused easily, drug sensitive report time is greatly shortened to only 4-6 h.
Description
Technical field
The invention belongs to technical field of microbial detection, in particular it relates to a kind of TTC for drug sensitive experiment is aobvious
Color culture medium and a kind of susceptibility kit and preparation method thereof and drug sensitive experiment method.
Background technology
Different bacteriums can occur different reactions to different antibiotic, and for a certain antibiotic sensitive, then this resists
Raw element can suppress the growth of bacterium, breeding, and then for therapeutic choice antibiotic medicine provides reference, we claim this process
Be drug sensitive experiment.
Whether traditional drug sensitive experiment room judges bacterium to a certain antibiotic sensitive by than turbid principle, it is impossible to intuitively
Observation drug sensitive experiment result.
The content of the invention
In order to overcome above technical problem, the present invention to provide a kind of TTC chromogenic culture mediums and one kind for drug sensitive experiment
Susceptibility kit and preparation method thereof and drug sensitive experiment method so that bacterium can intuitively show to the susceptibility results of medicine.
In order to reach object above, the present invention is adopted the following technical scheme that：A kind of TTC for drug sensitive experiment develops the color and cultivates
Base, including brain heart infusion broth（Abbreviation BHI meat soups）, acid protein hydrolysats, three tetrazoles of soluble starch and 2,3,5- chlorinations
（Abbreviation TTC）.Bacterium can produce succinate dehydrogenase, succinate dehydrogenase energy reductase 12,3,5- chlorinations during growth metabolism
Three tetrazoles（Also it is called TTC）, generate red San Ben formazans.In antimicrobial susceptibility experiment, if the growth and breeding of bacterium
Being suppressed by antibiotic medicine, succinate dehydrogenase would not be produced, entering without there is color change, you can judge bacterium to this
Antibiotic medicine is sensitive；And when antibiotic medicine does not have inhibitory action to the growth and breeding of bacterium, the growth and breeding meeting of bacterium
Succinate dehydrogenase is produced, succinate dehydrogenase and then TTC San Ben formazans is reduced into, so as to red change is presented, you can sentence
Disconnected bacterium is insensitive to the antibiotic medicine.Using technical scheme, drug sensitive experiment result can be intuitively observed, can be real
Existing intuitive judgment, easily observation, is less prone to deviation.
In any of the above-described scheme preferably, the TTC chromogenic culture mediums, based on percentage（Mass/volume ×
100%）, including TTC described in 0.01 ~ 0.03%.Traditional use judges drug sensitive experiment result than turbid principle, generally requires 18 ~ 24
Hour could correctly read susceptibility results of the bacterium to medicine, and time-consuming；And TTC chromogenic culture mediums of the invention are used, only
Need can obtain sensitivity experiments result of the bacterium to certain antibiotic medicine within 4 ~ 6 hours, greatly shorten susceptibility report
Time.
In any of the above-described scheme preferably, the TTC chromogenic culture mediums, based on percentage, including 0.015 ~
TTC described in 0.025%.
In any of the above-described scheme preferably, the TTC chromogenic culture mediums, based on percentage, including 0.018 ~
TTC described in 0.02%.
In any of the above-described scheme preferably, the TTC chromogenic culture mediums, based on percentage, including 2 ~ 5%BHI meat
Soup.BHI meat soups are nutritious, comprehensive, can provide nutrition for the breeding of the normal growth of bacterium.
In any of the above-described scheme preferably, the TTC chromogenic culture mediums, based on percentage, including 1 ~ 3% sour water
Solution albumen.Acid protein hydrolysats can provide the nutriments such as the nitrogen source for being easy to utilize for bacterial growth.Sour water solution used in this programme
Albumen is purchased from Chinese medicines group.
In any of the above-described scheme preferably, the TTC chromogenic culture mediums, based on percentage, including 0.05 ~ 2% can
Soluble starch.Soluble starch can provide the nutriments such as carbon source for bacterial growth.Soluble starch used is purchased from this programme
Chinese medicines group.
In any of the above-described scheme preferably, based on percentage, the TTC chromogenic culture mediums are by following component structure
Into：
BHI 2~5%
Acid protein hydrolysats 1 ~ 3%
Soluble starch 0.05 ~ 2%
TTC 0.01~0.03%
Purifying water surplus.
In any of the above-described scheme preferably, the TTC chromogenic culture mediums, based on percentage, by following component structure
Into：
BHI 2~5%
Acid protein hydrolysats 1 ~ 3%
Soluble starch 0.05 ~ 2%
TTC 0.015~0.025%
Purifying water surplus.
In any of the above-described scheme preferably, the TTC chromogenic culture mediums, based on percentage, by following component structure
Into：
BHI 2~5%
Acid protein hydrolysats 1 ~ 3%
Soluble starch 0.05 ~ 2%
TTC 0.018~0.02%
Purifying water surplus.
The TTC chromogenic culture mediums that the present invention is provided are used for drug sensitive experiment, can intuitively observe drug sensitive experiment result, easily see
Examine, be less prone to deviation, and susceptibility report time is greatly shortened, and is only needed 4 ~ 6 hours.
Second aspect, the present invention provides a kind of susceptibility kit, including kit of parts and susceptibility plate part, the reagent
Box part includes three tetrazole chromogenic culture mediums of 2,3,5- chlorinations（Abbreviation TTC chromogenic culture mediums）.
Preferably, the TTC chromogenic culture mediums that the TTC chromogenic culture mediums are provided for first aspect present invention.
In any of the above-described scheme preferably, the drug sensitive plate includes one or more susceptibility holes.
In any of the above-described scheme preferably, each described susceptibility hole includes biochemical reagents and antibiotic.
In any of the above-described scheme preferably, the biochemical reagents include BHI, acid protein hydrolysats, soluble starch, calcium from
Son, magnesium ion, comprising these biochemical reagents content with ratio phase in the TTC chromogenic culture mediums provided in this programme
Together.
In any of the above-described scheme preferably, each described susceptibility hole includes the antibiotic of 0.25 ~ 256 mcg/ml.
Preferably, for the bacterium that growth is difficult in any of the above-described scheme, addition growth factor, the growth may be selected
Factor Source is in serum and/or blood meal.Preferably to carry out drug sensitive experiment, more accurately drug sensitive experiment result is obtained.
In any of the above-described scheme preferably, each described susceptibility hole is also including growth factor that percentage is 37%.
The susceptibility kit that the present invention is provided, can intuitively observe drug sensitive experiment result, and easily observation, is less prone to deviation,
And susceptibility report time is greatly shortened, only need 4 ~ 6 hours.
Third aspect present invention provides a kind of preparation method of susceptibility kit, including prepares susceptibility plate part, also includes
Prepare TTC chromogenic culture mediums.
Preferably, the TTC chromogenic culture mediums that the TTC chromogenic culture mediums are provided for first aspect present invention.
In any of the above-described scheme preferably, the preparation method of the TTC chromogenic culture mediums includes：Each raw material is weighed to be dissolved in
Purified water, and be diluted with purified water, then packed, sterilized.
In any of the above-described scheme preferably, the sterilizing uses irradiation sterilization.
In any of the above-described scheme preferably, the irradiation dose of the irradiation sterilization is 8kGy.
In any of the above-described scheme preferably, the preparation method of the susceptibility kit, the susceptibility kit is this hair
The susceptibility kit that bright second aspect is provided.
In any of the above-described scheme preferably, the preparation method of the susceptibility plate part is comprised the following steps：
A. weigh each raw material and be dissolved in purified water；
B. it is diluted with purified water；
C. the solution after dilution is added into biochemical cup, and is dried, packs, and sterilized.
In any of the above-described scheme preferably, the sterilizing uses irradiation sterilization.
In any of the above-described scheme preferably, the irradiation dose of the irradiation sterilization is 8kGy.
In any of the above-described scheme preferably, the preparation method of the susceptibility kit comprises the following steps：
I. susceptibility plate part is prepared；
Ii. TTC chromogenic culture mediums are prepared；
Iii. susceptibility plate part and TTC chromogenic culture mediums are sterilized simultaneously.
In any of the above-described scheme preferably, the sterilizing uses irradiation sterilization.
In any of the above-described scheme preferably, the irradiation dose of the irradiation sterilization is 8kGy.
In any of the above-described scheme preferably, the preparation of the susceptibility plate part includes：Weigh each raw material and be dissolved in purifying
Water；Then it is diluted with purified water；Then the solution after dilution is added into biochemical cup, and is dried, packs.
The preparation method of the susceptibility kit that the present invention is provided, when the susceptibility kit for preparing is used for drug sensitive experiment,
Drug sensitive experiment result can be intuitively observed, easily observation, is less prone to deviation, and susceptibility report time is greatly shortened, only need 4 ~ 6 are small
When.
A kind of fourth aspect present invention, there is provided drug sensitive experiment method, including bacteria suspension is diluted with culture medium and adds medicine
Quick hole, and cultivated, wherein, the TTC chromogenic culture mediums that the culture medium is provided for first aspect present invention.
Preferably, the drug sensitive experiment method is comprised the following steps：
A, prepare the TTC chromogenic culture mediums；
B, with the TTC chromogenic culture mediums dilute bacteria suspension, be configured to TTC colour developing Analysis of Drug Susceptibility liquid；
C, Analysis of Drug Susceptibility liquid that the TTC develops the color are added in susceptibility hole, and then sealing is cultivated, observes result.
In any of the above-described scheme preferably, in the susceptibility plate part that the susceptibility hole is related to for second aspect present invention
One or more susceptibility holes.
In any of the above-described scheme preferably, the preparation method ginseng of the TTC chromogenic culture mediums and the susceptibility plate part
The TTC chromogenic culture medium preparation methods of third aspect present invention offer are provided.
In any of the above-described scheme preferably, the bacteria suspension uses the bacterium of fresh passage, i.e. passed within 24 hours
Generation.
In any of the above-described scheme preferably, the concentration of the bacteria suspension is 0.5 maxwell unit.
In any of the above-described scheme preferably, 100 microlitres of TTC colour developings Analysis of Drug Susceptibility are added in each described susceptibility hole
Liquid.
In any of the above-described scheme preferably, the culture is carried out in 35 ~ 37 DEG C of incubators.
In any of the above-described scheme preferably, the observation result is carried out after cultivating 4 ~ 6 hours.
The drug sensitive experiment method provided using the present invention, can intuitively observe drug sensitive experiment result, and easily observation, is less prone to
Deviation, and susceptibility report time greatly shortens, and only needs 4 ~ 6 hours.
Specific embodiment
In order to more correctly understand present disclosure, present disclosure is done into one with reference to specific embodiment
Step explanation.BHI meat soups used in following examples are bought from OXOID, and its specific composition is：Beef heart infusion agar 30%, albumen
Peptone 1.5%, sodium chloride 1%；, from parent company of Chinese Medicine group, TTC purchases are certainly for acid protein hydrolysats used and soluble starch purchase
Sigma companies.
Embodiment 1
A kind of TTC chromogenic culture mediums for drug sensitive experiment, including brain heart infusion broth（Abbreviation BHI meat soups）, acid protein hydrolysats,
Three tetrazoles of soluble starch and 2,3,5- chlorinations（Abbreviation TTC）, specifically, according to percentage meter, this is used for susceptibility
The TTC chromogenic culture mediums of experiment are made up of following component：
BHI meat soups 5%
Acid protein hydrolysats 3%
Soluble starch 2%
TTC 0.03%。
Specifically, its quality and volume are as follows：
BHI meat soups 5mg
Acid protein hydrolysats 3mg
Soluble starch 2mg
TTC 0.03mg
Purified water 100ml.
Bacterium can produce succinate dehydrogenase, succinate dehydrogenase energy reductase 12,3,5- chlorinations three during growth metabolism
This tetrazole（Also it is called TTC）, generate red San Ben formazans.In antimicrobial susceptibility experiment, if the growth and breeding quilt of bacterium
Antibiotic medicine suppresses, and would not produce succinate dehydrogenase, enters without there is color change, you can judge that bacterium is anti-to this
Raw element medicaments insensitive；And when antibiotic medicine does not have inhibitory action to the growth and breeding of bacterium, the growth and breeding of bacterium can be produced
Unripe amber acidohydrogenase, succinate dehydrogenase and then is reduced into San Ben formazans by TTC, so as to red change is presented, you can judge
Bacterium is insensitive to the antibiotic medicine.Using technical scheme, drug sensitive experiment result can be intuitively observed, be capable of achieving
Intuitive judgment, easily observation, is less prone to deviation.Using TTC principles, it is only necessary to bacterium can be obtained within 4 ~ 6 hours to certain antibiosis
The sensitivity experiments result of plain medicine, greatly shortens the time of susceptibility report.
On this basis, a kind of susceptibility kit is prepared for, the susceptibility kit includes kit of parts and susceptibility plate portion
Point, the kit of parts includes above-mentioned 2,3,5- three tetrazole chromogenic culture mediums of chlorination（Abbreviation TTC chromogenic culture mediums）, its
In, drug sensitive plate includes one or more susceptibility holes, and specifically, in the present embodiment, drug sensitive plate is 96 orifice plates.In each susceptibility hole
Including antibiotic and biochemical reagents, specifically, biochemical reagents refer to BHI meat soups, acid protein hydrolysats, soluble starch, calcium ion,
Magnesium ion etc., this can be matched according to existing technology, and in the present embodiment, biochemical reagents percentage is 0.5-15%,
The amount of antibiotic is the mcg/ml of 0.5 mcg/ml -256.
The preparation method of above susceptibility kit, including susceptibility plate part is prepared, also including preparing TTC chromogenic culture mediums.
Specifically, the preparation method of the susceptibility kit is as follows：
The preparation method of TTC chromogenic culture mediums：Weigh each raw material of the above and be dissolved in appropriate purified water, for the purpose of dissolving, and with pure
Change water to be diluted, then packed, sterilized；Wherein, the sterilizing uses irradiation sterilization, and irradiation dose is 8kGy.
The preparation of susceptibility plate part is comprised the following steps successively：
A. weigh each raw material and be dissolved in purified water：Weigh biochemical reagents（BHI meat soups, acid protein hydrolysats, soluble starch）With it is anti-
Raw element is dissolved in purified water；
B. it is diluted with purified water：The solution that biochemical reagents and antibiotic will be dissolved with is diluted with purified water；
C. the solution after dilution is added into biochemical cup, and is dried, packs, and sterilized：Solution after dilution is added
In the biochemical hole of 96 orifice plates, as susceptibility hole, 30 microlitres of solution after dilution is added in each hole, 96 orifice plates are then placed in 45
Dried at DEG C, after drying, 96 orifice plates are packed and sterilization treatment is carried out, wherein, sterilizing uses irradiation sterilization, irradiates
Dosage is 8kGy, and sterilization time is 4-6 hours.
And then, the present embodiment provides a kind of drug sensitive experiment method, in turn includes the following steps：
First, with the bacterium of fresh passage（In 24 hours）Configuration concentration is the bacteria suspension of 0.5 Maxwell concentration；
Secondly, bacteria suspension is diluted into 100 with the TTC chromogenic culture mediums to be simultaneously well mixed, is configured to TTC colour developing Analysis of Drug Susceptibility
Liquid；
Then, in the drug sensitive plate（96 orifice plates）Susceptibility hole（The hole of 96 orifice plates）It is middle to add 100 microlitres of TTC colour developings susceptibility
Analysis liquid, covers lid；
Finally, drug sensitive plate is placed in 35-37 DEG C of incubator and is cultivated, and observe result.
Certainly, when drug sensitive experiment is carried out, TTC chromogenic culture mediums can also be Extemporaneous, and this when,
The preparation of TTC chromogenic culture mediums and the preparation part sequencing of bacteria suspension.
In the present embodiment, the bacteria suspension is prepared using EHEC ATCC25922, using TTC chromogenic culture medium generations
Bacteria suspension is diluted for common susceptibility nutrient solution, antibiotic selects Cefepime, Cefepime is added in each susceptibility hole
0.25 mcg/ml, according to the present embodiment, cultivates 4 hours, you can substantially observe color change in incubator.
And use commonsense method of the prior art（For example, formula is：BHI meat soups 5%, acid protein hydrolysats 3%, solubility
Starch 2%, using than turbid principle）, it is necessary to 16 hours could substantially observe drug sensitive experiment result.
Embodiment 2
A kind of TTC chromogenic culture mediums for drug sensitive experiment, as different from Example 1, for the bacterium of difficult growth, often
Also include growth factor in individual susceptibility hole, the addition of growth factor can be added according to prior art, and such as percentage is
30%.Growth factor can be added by adding blood meal, serum, preferably to carry out drug sensitive experiment, obtain more accurately susceptibility
Experimental result.
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.1
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 2%
Acid protein hydrolysats 1%
Soluble starch 0.05%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.2
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 2%
Acid protein hydrolysats 1%
Soluble starch 0.05%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.3
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 3%
Acid protein hydrolysats 2%
Soluble starch 0.15%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.4
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 4%
Acid protein hydrolysats 3%
Soluble starch 0.1%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.5
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 3%
Acid protein hydrolysats 3%
Soluble starch 1.5%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.6
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 3%
Acid protein hydrolysats 3%
Soluble starch 1%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.7
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 3.2%
Acid protein hydrolysats 1.5%
Soluble starch 0.2%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.8
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 3.5%
Acid protein hydrolysats 1.25%
Soluble starch 0.15%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.9
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 3.7%
Acid protein hydrolysats 1.75%
Soluble starch 0.15%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.10
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 3.9%
Acid protein hydrolysats 1.75%
Soluble starch 0.15%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.11
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 2.7%
Acid protein hydrolysats 2.5%
Soluble starch 0.75%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 3.12
A kind of TTC chromogenic culture mediums for drug sensitive experiment, it is unlike the embodiments above, according to percentage meter, should
TTC chromogenic culture mediums for drug sensitive experiment are made up of following component：
BHI meat soups 3.2%
Acid protein hydrolysats 1.5%
Soluble starch 0.2%
TTC 0.03%。
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
This group of embodiment be used to illustrate in TTC chromogenic culture mediums, and BHI meat soups, acid protein hydrolysats and soluble starch can be with
It is adjusted according to actual needs, specifically chosen scope（Percentage）For：BHI meat soups 2 ~ 5%, acid protein hydrolysats 1 ~ 3%, can
Soluble starch 0.05 ~ 2%.
Embodiment 4.1
A kind of TTC chromogenic culture mediums for drug sensitive experiment, unlike the embodiments above, the percentage content of TTC
It is 0.01%.
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 4.2
A kind of TTC chromogenic culture mediums for drug sensitive experiment, unlike the embodiments above, the percentage content of TTC
It is 0.015%.
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 4.3
A kind of TTC chromogenic culture mediums for drug sensitive experiment, unlike the embodiments above, the percentage content of TTC
It is 0.018%.
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 4.4
A kind of TTC chromogenic culture mediums for drug sensitive experiment, unlike the embodiments above, the percentage content of TTC
It is 0.02%.
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 4.5
A kind of TTC chromogenic culture mediums for drug sensitive experiment, unlike the embodiments above, the percentage content of TTC
It is 0.024%.
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 4.6
A kind of TTC chromogenic culture mediums for drug sensitive experiment, unlike the embodiments above, the percentage content of TTC
It is 0.028%.
The preparation method of TTC chromogenic culture mediums, therefore, it is changed accordingly.
Embodiment 5
A kind of preparation method of susceptibility kit, it is unlike the embodiments above, for temperature-sensitive antibiotic, in susceptibility plate part
During preparation, its drying can be carried out using freeze-drying, the specific method of freeze-drying may refer to prior art or
Patent of the person before us.
Embodiment 6.1
A kind of drug sensitive experiment kit, unlike the embodiments above, in the present embodiment, antibiotic selects gentamicin.Medicine
The preparation method of quick kit, therefore, it is changed accordingly.
When carrying out drug sensitive experiment, cultivated 5 hours in incubator, you can substantially observe color change.
And commonsense method of the prior art is used, it is necessary to 18 hours could substantially observe drug sensitive experiment result.
Embodiment 6.2
A kind of drug sensitive experiment kit, unlike the embodiments above, in the present embodiment, antibiotic selects azithromycin.Medicine
The preparation method of quick kit, therefore, it is changed accordingly.
When carrying out drug sensitive experiment, cultivated 4 hours in incubator, you can substantially observe color change.
And commonsense method of the prior art is used, it is necessary to 18 hours could substantially observe drug sensitive experiment result.
Embodiment 6.3
A kind of drug sensitive experiment kit, unlike the embodiments above, in the present embodiment, antibiotic selects Cefepime.Medicine
The preparation method of quick kit, therefore, it is changed accordingly.
When carrying out drug sensitive experiment, bacteria suspension is prepared using staphylococcus aureus ATCC25923, according to this implementation
Example, cultivates 5 hours, you can substantially observe color change in incubator.
And commonsense method of the prior art is used, it is necessary to 18 hours could substantially observe drug sensitive experiment result.
Embodiment 6.4
A kind of drug sensitive experiment kit, unlike the embodiments above, in the present embodiment, antibiotic selects gentamicin.Medicine
The preparation method of quick kit, therefore, it is changed accordingly.
When carrying out drug sensitive experiment, bacteria suspension is prepared using staphylococcus aureus ATCC25923, according to this implementation
Example, cultivates 6 hours, you can substantially observe color change in incubator.
And commonsense method of the prior art is used, it is necessary to 20 hours could substantially observe drug sensitive experiment result.
Embodiment 6.5
A kind of drug sensitive experiment kit, unlike the embodiments above, in the present embodiment, antibiotic selects azithromycin.Medicine
The preparation method of quick kit, therefore, it is changed accordingly.
When carrying out drug sensitive experiment, bacteria suspension is prepared using staphylococcus aureus ATCC25923, according to this implementation
Example, cultivates 5 hours, you can substantially observe color change in incubator.
And commonsense method of the prior art is used, it is necessary to 18 hours could substantially observe drug sensitive experiment result.
Embodiment 7.1
A kind of susceptibility kit, unlike the embodiments above, in the susceptibility hole of kit, the content of antibiotic is 0.5 micro-
Grams per milliliter.The preparation method of susceptibility kit, therefore, it is changed accordingly.
Embodiment 7.2
A kind of susceptibility kit, it is unlike the embodiments above, in the susceptibility hole of kit, the content of antibiotic for 1 microgram/
Milliliter.The preparation method of susceptibility kit, therefore, it is changed accordingly.
Embodiment 7.3
A kind of susceptibility kit, it is unlike the embodiments above, in the susceptibility hole of kit, the content of antibiotic for 2 micrograms/
Milliliter.The preparation method of susceptibility kit, therefore, it is changed accordingly.
Embodiment 7.4
A kind of susceptibility kit, it is unlike the embodiments above, in the susceptibility hole of kit, the content of antibiotic for 4 micrograms/
Milliliter.The preparation method of susceptibility kit, therefore, it is changed accordingly.
Embodiment 7.5
A kind of susceptibility kit, unlike the embodiments above, in the susceptibility hole of kit, the content of antibiotic is 16 micro-
Grams per milliliter.The preparation method of susceptibility kit, therefore, it is changed accordingly.
Embodiment 7.6
A kind of susceptibility kit, unlike the embodiments above, in the susceptibility hole of kit, the content of antibiotic is 32 micro-
Grams per milliliter.The preparation method of susceptibility kit, therefore, it is changed accordingly.
Embodiment 7.7
A kind of susceptibility kit, unlike the embodiments above, in the susceptibility hole of kit, the content of antibiotic is 64 micro-
Grams per milliliter.The preparation method of susceptibility kit, therefore, it is changed accordingly.
Embodiment 7.8
A kind of susceptibility kit, unlike the embodiments above, in the susceptibility hole of kit, the content of antibiotic is 128 micro-
Grams per milliliter.The preparation method of susceptibility kit, therefore, it is changed accordingly.
Embodiment 7.9
A kind of susceptibility kit, unlike the embodiments above, in the susceptibility hole of kit, the content of antibiotic is 256 micro-
Grams per milliliter.The preparation method of susceptibility kit, therefore, it is changed accordingly.
No matter this group of embodiment is for illustrating that the concentration of antibiotic is how many, using TTC chromogenic culture mediums of the invention, medicine
Quick kit and drug sensitive experiment method, can in a short time substantially, be visually observed that because bacterium can not be pressed down by antibiotic
TTC is reduced into the San Ben formazans of red and culture medium is changed colour during system growth, significantly shorten drug sensitive experiment report time,
And, easily determine, it is less prone to deviation.
Embodiment 8
A kind of preparation method of susceptibility kit, it is unlike the embodiments above, in the present embodiment, the preparation of susceptibility kit
Method includes successively：
First, susceptibility plate part is prepared：Weigh each raw material and be dissolved in purified water：Weigh biochemical reagents（BHI meat soups, sour water solution egg
In vain, soluble starch）And antibiotic, or, being dissolved in purified water with growth factor；It is diluted with purified water：Biochemistry will be dissolved with
The solution of reagent and antibiotic is diluted with purified water；Solution after dilution is added into biochemical cup, and is dried, packed；
Secondly, TTC chromogenic culture mediums are prepared：Weigh each raw material of the above and be dissolved in appropriate purified water, for the purpose of dissolving, and with purifying
Water is diluted, dispenses；
Finally, susceptibility plate part and TTC chromogenic culture mediums are sterilized simultaneously, other conditions and a kind of upper preparation method phase
Together.This susceptibility reagent box preparation method is particularly suited for the feelings that susceptibility plate part and kit of parts are preserved in same packaging
Condition.
In the present invention, the preparation method of susceptibility kit can be selected according to actual conditions, susceptibility plate part and examination
Agent box part can each pack, sterilize；Can also be after both each pack while being sterilized.
It should be noted that various embodiments above is merely to illustrate technical scheme, rather than its limitations；Although
The present invention has been described in detail with reference to foregoing embodiments, it will be understood by those within the art that；It is still
Technical scheme described in foregoing embodiments can be modified, or which part or all technical characteristic are carried out
Equivalent；And these modifications or replacement, the essence of appropriate technical solution is departed from various embodiments of the present invention technical side
The scope of case.
Claims (10)
1. a kind of TTC chromogenic culture mediums for drug sensitive experiment, including brain heart infusion broth, acid protein hydrolysats, soluble starch,
It is characterized in that：Also include three tetrazoles of 2,3,5- chlorinations.
2. TTC chromogenic culture mediums as claimed in claim 1, it is characterised in that：The TTC chromogenic culture mediums, by concentration percentage
Than meter, including TTC described in 0.01-0.03%.
3. a kind of susceptibility kit, including kit of parts and susceptibility plate part, it is characterised in that：The kit of parts includes
TTC chromogenic culture mediums.
4. susceptibility kit as claimed in claim 3, it is characterised in that：The TTC chromogenic culture mediums are in claim 1-2
TTC chromogenic culture mediums described in any one.
5. susceptibility kit as claimed in claim 4, it is characterised in that：The drug sensitive plate includes one or more susceptibility holes.
6. susceptibility kit as claimed in claim 5, it is characterised in that：Each described susceptibility hole includes biochemical reagents and antibiosis
Element.
7. susceptibility kit as claimed in claim 6, it is characterised in that：The biochemical reagents include BHI, acid protein hydrolysats,
Soluble starch, calcium ion, magnesium ion.
8. a kind of preparation method of susceptibility kit, including preparation susceptibility plate part, it is characterised in that：Also include that preparing TTC shows
Color culture medium.
9. the preparation method of susceptibility kit as claimed in claim 8, it is characterised in that：The TTC chromogenic culture mediums are power
Profit requires the TTC chromogenic culture mediums any one of 1-2.
10. a kind of drug sensitive experiment method, including bacteria suspension is diluted with culture medium and adds susceptibility hole, and cultivated, it is special
Levy and be, the culture medium is the TTC chromogenic culture mediums any one of claim 1-2.
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| CN201610448317.0A CN106701892A (en) | 2016-06-21 | 2016-06-21 | TTC (2,3,5-triphenyltetrazolium chloride) chromogenic culture medium for drug sensitivity tests, drug sensitivity kit, preparation method of drug sensitivity kit and drug sensitivity test method |
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| CN201610448317.0A CN106701892A (en) | 2016-06-21 | 2016-06-21 | TTC (2,3,5-triphenyltetrazolium chloride) chromogenic culture medium for drug sensitivity tests, drug sensitivity kit, preparation method of drug sensitivity kit and drug sensitivity test method |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111088318A (en) * | 2020-01-03 | 2020-05-01 | 西北农林科技大学 | Method for detecting bacterial drug sensitivity by TTC reduction reaction |
| CN111118104A (en) * | 2018-10-30 | 2020-05-08 | 深圳市帝迈生物技术有限公司 | Culture medium and preparation method thereof, kit, detection device and detection method |
| CN112378902A (en) * | 2020-11-03 | 2021-02-19 | 华中农业大学 | Method for rapidly evaluating disinfection effect of antibacterial disinfectant |
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|---|---|---|---|---|
| CN1110988A (en) * | 1994-04-28 | 1995-11-01 | 钟国华 | Quick medicine-sensitive subtratum |
| CN102888442A (en) * | 2011-07-20 | 2013-01-23 | 农高惠 | Kit for rapid culture, identification and drug sensitivity of gastric helicobacter pylori and inspection method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1110988A (en) * | 1994-04-28 | 1995-11-01 | 钟国华 | Quick medicine-sensitive subtratum |
| CN102888442A (en) * | 2011-07-20 | 2013-01-23 | 农高惠 | Kit for rapid culture, identification and drug sensitivity of gastric helicobacter pylori and inspection method thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111118104A (en) * | 2018-10-30 | 2020-05-08 | 深圳市帝迈生物技术有限公司 | Culture medium and preparation method thereof, kit, detection device and detection method |
| CN111088318A (en) * | 2020-01-03 | 2020-05-01 | 西北农林科技大学 | Method for detecting bacterial drug sensitivity by TTC reduction reaction |
| CN112378902A (en) * | 2020-11-03 | 2021-02-19 | 华中农业大学 | Method for rapidly evaluating disinfection effect of antibacterial disinfectant |
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