CN106119336A - A kind of improvement glucose peptone water culture medium being used for carrying out VP test - Google Patents
A kind of improvement glucose peptone water culture medium being used for carrying out VP test Download PDFInfo
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- CN106119336A CN106119336A CN201610507987.5A CN201610507987A CN106119336A CN 106119336 A CN106119336 A CN 106119336A CN 201610507987 A CN201610507987 A CN 201610507987A CN 106119336 A CN106119336 A CN 106119336A
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- culture medium
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- peptone water
- glucose
- creatinine
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/10—Enterobacteria
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/045—Culture media therefor
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- Proteomics, Peptides & Aminoacids (AREA)
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Abstract
The present invention provides a kind of improvement glucose peptone water culture medium for carrying out VP test, and described culture medium is made up of each component of following quality volumn concentration: 1% common glucose glycoprotein peptone water, the creatinine of 1%.The culture medium of the application present invention carries out VP test and can speed up bacterial growth, thus reduces VP and test required time, makes the time foreshorten to 46 hours.
Description
Technical field
The present invention relates to a kind of improvement glucose peptone water culture medium for carrying out VP test.
Background technology
The reaction principle of VP test is: some antibacterial energy decomposition glucose produces acetone acid, is further taken off by acetone acid
Hydroxyl becomes acetyl methyl carbinol, is oxidized to diacetyl, diacetyl and albumen in culture medium by the oxygen in air in alkaline environment
In peptone, guanidine radicals contained by arginine reacts and generates red compound.It is mainly used in the discriminating of Enterobacter bacteria.Sha Lei
The VP reacting positive such as Salmonella, enterobacter cloacae, the VP reaction negative such as escherichia coli, Salmonella, shigella.
Existing VP test method is:
According to laboratory usage amount, prepare 300-500ml 1% common glucose glycoprotein peptone water, take 2ml and be sub-packed in 12 × 150mm
In test tube, add a cover silica gel plug, 115 DEG C of 10 pounds of 15 minutes autoclavings.After reagent cooling, filling plastic bag tying, 4 DEG C of refrigerators are protected
Deposit.
Reaction reagent:
Solution A: 6% alpha-Naphthol alcoholic solution；
Second liquid: 40% potassium hydroxide.
Test method: picking pure culture inoculation medium.In 35 DEG C of cultivations, cultivate 18～24h.
Result is observed: after cultivation, adds each 3 of Barritt VP reagent liquid A and liquid B, and observed result after shaking, in number
For the positive when taking on a red color in minute, as redfree occurs being feminine gender.
Quality-control strains: positive strain: ATCC700603 Klebsiella Pneumoniae；
Negative strain: ATCC25922 escherichia coli.
The above-mentioned VP test traditional technique in measuring time is long, need to can check result, time-consumingly with after culture medium 18-24 hour
Long.
Summary of the invention
The invention provides a kind of improvement glucose peptone water culture medium for carrying out VP test, it is possible to accelerate antibacterial
Growth, thus reduce VP and test required time.
First purpose of the present invention is to provide a kind of improvement glucose peptone water culture medium for carrying out VP test,
Described culture medium is made up of each component of following weight/mass percentage composition: 1% common glucose glycoprotein peptone water, the creatinine of 1%.Mass body
The unit of long-pending percentage ratio is g/ml.
Second object of the present invention is to provide the system of the improvement glucose peptone water culture medium for carrying out VP test
Preparation Method, adds the creatinine of 1%, autoclaving and get final product in 1% common glucose glycoprotein peptone water.
Third object of the present invention is to provide the improvement glucose peptone water culture medium for carrying out VP test at VP
Application in test.
Fourth object of the present invention is to provide a kind of VP test method, and step is as follows:
(1) culture medium is prepared: in 1% common glucose glycoprotein peptone water, add the creatinine of 1%, autoclaving；
(2) preparation reaction reagent: solution A: 6% alpha-Naphthol alcoholic solution；
Second liquid: 40% potassium hydroxide；
(3) test: culture medium prepared by picking pure culture inoculation step (1) to be detected, in 35 DEG C of cultivations, cultivate 4
～6h；
(4) result is observed: after cultivation, adds each 3 of liquid A and liquid B, and observed result after shaking, when taking on a red color in several minutes
Occur being feminine gender for the positive, such as redfree.
The culture medium of the application present invention carries out VP test and can speed up bacterial growth, thus reduces VP and test required time,
The time is made to foreshorten to 4-6 hour.
Detailed description of the invention
Below example facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiment
Method, if no special instructions, is conventional method.Test material used in following embodiment, if no special instructions, is city
Sell.If no special instructions, in the present invention, " % " is quality percent by volume.
Embodiment 1
The present invention provides a kind of improvement glucose peptone water culture medium for carrying out VP test, and formula is:
1% common glucose glycoprotein peptone water, the creatinine of 1%.
The preparation method of a kind of improvement glucose peptone water culture medium for carrying out VP test of the present invention is:
According to laboratory usage amount, prepare 1% common glucose glycoprotein peptone water 300～500ml, add the creatinine of 1%, for accelerating
Reacting, quick and precisely identify, the applicant separately adds 1% creatinine in common 1% glucose peptone water basal medium.Take
0.5ml adds in 2ml cillin bottle, sealing.115 DEG C of 10 pounds of 15 minutes autoclavings.After reagent cooling, room temperature preservation.
The improved culture medium of the application present invention carries out VP test method particularly includes:
Reaction reagent:
Solution A: 6% alpha-Naphthol alcoholic solution；
Second liquid: 40% potassium hydroxide.
Test method: the improved culture medium of the pure culture inoculation present invention that picking is to be detected, in 35 DEG C of cultivations, cultivates 4
～6h.
Result is observed: after cultivation, adds each 3 of Barritt VP reagent liquid A and liquid B, and observed result after shaking, in number
For the positive when taking on a red color in minute, as redfree occurs being feminine gender.
Quality-control strains: positive strain: ATCC700603 Klebsiella Pneumoniae；
Negative strain: ATCC25922 escherichia coli.
Apply different culture medium, identical VP experimental condition and method respectively, different bacterial strains to be checked are tested, training
The time of supporting is as shown in table 1, and testing result is the most identical.
Table 1
Finally it is noted that the foregoing is only the preferred embodiments of the present invention, it is not limited to the present invention, although
Being described in detail the present invention with reference to previous embodiment, for a person skilled in the art, it still can be right
Technical scheme described in foregoing embodiments is modified, or wherein portion of techniques feature is carried out equivalent.All
Within the spirit and principles in the present invention, any modification, equivalent substitution and improvement etc. made, should be included in the protection of the present invention
Within the scope of.
Claims (4)
1. one kind is used for carrying out the improvement glucose peptone water culture medium of VP test, it is characterised in that: described culture medium by with
Each component composition of lower quality volumn concentration: 1% common glucose glycoprotein peptone water, the creatinine of 1%.
2. being used for described in claim 1 carries out the preparation method improveing glucose peptone water culture medium of VP test, and it is special
Levy and be: in 1% common glucose glycoprotein peptone water, add the creatinine of 1%, autoclaving and get final product.
3. being used for described in claim 1 carries out improvement glucose peptone water culture medium the answering in VP tests of VP test
With.
4. a VP test method, it is characterised in that: step is as follows:
(1) culture medium is prepared: in 1% common glucose glycoprotein peptone water, add the creatinine of 1%, autoclaving；Described percentage ratio is
Quality percent by volume；
(2) preparation reaction reagent: solution A: 6% alpha-Naphthol alcoholic solution；
Second liquid: 40% potassium hydroxide；
(3) test: culture medium prepared by picking pure culture inoculation step (1) to be detected, in 35 DEG C of cultivations, cultivate 4
～6h；
(4) result is observed: after cultivation, adds each 3 of liquid A and liquid B, and observed result after shaking, when taking on a red color in several minutes
Occur being feminine gender for the positive, such as redfree.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610507987.5A CN106119336A (en) | 2016-07-01 | 2016-07-01 | A kind of improvement glucose peptone water culture medium being used for carrying out VP test |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610507987.5A CN106119336A (en) | 2016-07-01 | 2016-07-01 | A kind of improvement glucose peptone water culture medium being used for carrying out VP test |
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|---|---|
| CN106119336A true CN106119336A (en) | 2016-11-16 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201610507987.5A Pending CN106119336A (en) | 2016-07-01 | 2016-07-01 | A kind of improvement glucose peptone water culture medium being used for carrying out VP test |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110551792A (en) * | 2019-08-20 | 2019-12-10 | 马玥 | Rapid V-P test method and kit used for same |
-
2016
- 2016-07-01 CN CN201610507987.5A patent/CN106119336A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 崔有宏等: "肌酐测定工具酶产生菌—烟草节杆菌02181的筛选及其产酶条件", 《甘肃科学学报》 * |
| 钟楠楠: "《微生物检验技术》", 31 December 2013 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110551792A (en) * | 2019-08-20 | 2019-12-10 | 马玥 | Rapid V-P test method and kit used for same |
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Application publication date: 20161116 |