CN104894039B - A kind of method and its special culture media of fast culture Pseudomonas aeruginosa - Google Patents
A kind of method and its special culture media of fast culture Pseudomonas aeruginosa Download PDFInfo
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- 241000589517 Pseudomonas aeruginosa Species 0.000 title claims abstract description 36
- 239000001963 growth medium Substances 0.000 title claims abstract description 28
- 238000000034 method Methods 0.000 title claims description 9
- 239000002609 medium Substances 0.000 claims abstract description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- WLJVXDMOQOGPHL-UHFFFAOYSA-N phenylacetic acid Chemical compound OC(=O)CC1=CC=CC=C1 WLJVXDMOQOGPHL-UHFFFAOYSA-N 0.000 claims description 6
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 4
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- 235000015097 nutrients Nutrition 0.000 description 4
- IGXWBGJHJZYPQS-SSDOTTSWSA-N D-Luciferin Chemical compound OC(=O)[C@H]1CSC(C=2SC3=CC=C(O)C=C3N=2)=N1 IGXWBGJHJZYPQS-SSDOTTSWSA-N 0.000 description 3
- CYCGRDQQIOGCKX-UHFFFAOYSA-N Dehydro-luciferin Natural products OC(=O)C1=CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 CYCGRDQQIOGCKX-UHFFFAOYSA-N 0.000 description 3
- BJGNCJDXODQBOB-UHFFFAOYSA-N Fivefly Luciferin Natural products OC(=O)C1CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 BJGNCJDXODQBOB-UHFFFAOYSA-N 0.000 description 3
- DDWFXDSYGUXRAY-UHFFFAOYSA-N Luciferin Natural products CCc1c(C)c(CC2NC(=O)C(=C2C=C)C)[nH]c1Cc3[nH]c4C(=C5/NC(CC(=O)O)C(C)C5CC(=O)O)CC(=O)c4c3C DDWFXDSYGUXRAY-UHFFFAOYSA-N 0.000 description 3
- 239000002054 inoculum Substances 0.000 description 3
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- 244000052769 pathogen Species 0.000 description 2
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- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 238000009423 ventilation Methods 0.000 description 2
- 241000244203 Caenorhabditis elegans Species 0.000 description 1
- 244000025254 Cannabis sativa Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
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- 206010040047 Sepsis Diseases 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
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- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical compound CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 description 1
- KCIDZIIHRGYJAE-YGFYJFDDSA-L dipotassium;[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate Chemical compound [K+].[K+].OC[C@H]1O[C@H](OP([O-])([O-])=O)[C@H](O)[C@@H](O)[C@H]1O KCIDZIIHRGYJAE-YGFYJFDDSA-L 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
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- 238000005457 optimization Methods 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 210000002640 perineum Anatomy 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- -1 phosphoric acid hydrogen Chemical class 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 1
- 235000008160 pyridoxine Nutrition 0.000 description 1
- 239000011677 pyridoxine Substances 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of culture medium that can promote Pseudomonas aeruginosa fast breeding.The culture medium is a kind of culture medium with preferable culture effect for groping to obtain by thousands of times by numerous components.The culture medium of the promotion Pseudomonas aeruginosa fast breeding of the present invention can significantly increase enhancing cell viability.The culture medium of the promotion bacteria fast reproduction of the present invention has the obvious effect for promoting bacterial growth, shortening incubation time compared with conventional medium.
Description
Technical field
The present invention relates to biological field, and in particular to a kind of method and its special culture media of fast culture Pseudomonas aeruginosa.
Background technology
Pseudomonas aeruginosa, also known as pseudomonas aeruginosa (scientific name Pseudomonas aeruginos), are a kind of Gram-negatives
Bacterium, aerobic, in long rod bacterium, only unidirectional motility.Green pus liver bacterium is a kind of conditioned pathogen, and external environment is fitted
Should be very capable, in wet condition can long-term surviving.People's normal skin especially moist position (such as oxter, perineum, ear
Road, respiratory tract and enteron aisle etc.) there is the presence of green pus liver bacterium, and complete skin and mucous membrane can be supported effectively as barrier
The invasion of imperial green pus liver bacterium, but the normal defense mechanisms of human body decline or sustain damage that (such as skin mucosa injury, granulocyte lacks
Weary, Hypoproteinemia, the chemicotherapy of tumour, prolonged application hormone and antibiotic, burn, infant's skin, umbilical cord, enteron aisle, always
Year people urinary system) then easily trigger the infection of green pus liver bacterium.
As the bacterium of other pseudomonas, Pseudomonas aeruginosa secretes a variety of pigments, including pyo (in green grass or young crops
Color), luciferin (being in fluorescent yellow) and pyorubin (being in blush).Pseudomonas culture medium P is just used as increasing green
The production of pyocin and pyorubin, and pseudomonas culture medium F is exactly the generation for strengthening luciferin.
Pseudomonas aeruginosa is characterized in it that grape smell such as pearlized profile and in vitro.Clinic confirms green pus bar
The method of bacterium is to be the generation of pyo and luciferin, and the ability grown in the environment of 42 DEG C.Pseudomonas aeruginosa is in bavin
Remain to grow in oil and aviation fuel, be more referred to as hydrogen carbon decomposer, Microbial Corrosion of Mild Steel can be triggered.It can produce a kind of dark
The gel mat of color, is typically misinterpreted as algae.
Pseudomonas aeruginosa is a kind of opportunistic infections cause of disease for making immunocompromised host, General Influence lung and the urinary tract, or is caused
Burn, wound and other blood infections, such as septicaemia.Although seldom having very much, Pseudomonas aeruginosa also results in pneumonia.In many lungs
Pointed out in inflammation research, Pseudomonas aeruginosa is the bacterium that one of which needs to completely cut off.Pyo is exactly the virulence factor of this bacterium,
And Caenorhabditis elegans can be made dead under oxidative stress, but the generation of pyo can be suppressed by having research to refer to salicylic acid.
10% is all led in the illness of hospital infection by Pseudomonas aeruginosa.The lung of cystic fibrosis patient be infect at first it is green
The a group of purulence bacillus.In the case where lacking proper treatment drinking water quality, it is also the one of which bacterium for being led to dermatitis.It is also to make
Into the most common bacterium of infection of burn.It is reported that nosocomial infection accounts for 10%~31.5% caused by Pseudomonas aeruginosa, occupy pathogen it
Head, domestic and international Burn Center is to finding that charrin's disease accounts for more than 50% in the clinical investigation of bacterium infection.Caused based on more than
Characteristic of disease, Pseudomonas aeruginosa are that one of microorganism must be examined in medical diagnosis on disease.Traditional detection method is according to, it is necessary to time of at least 3 days
Carry out bacterium colony culture detection.For the diagnosis of disease, there is a larger hysteresis quality.Therefore it provides one kind can be fast
The culture medium of speed culture Pseudomonas aeruginosa seems extremely important.
The content of the invention
The problem of it is an object of the invention to exist for above-mentioned various existing culture mediums, there is provided a kind of quick and culture sun
The property high Pseudomonas aeruginosa fluid nutrient medium of rate.
The invention provides following technical scheme：A kind of culture medium (1L) that can promote green pus bacteria fast reproduction：Benzene
Acetic acid 1.2g, casein peptone 5g, magnesium sulfate 0.08g, sodium chloride 2g, dipotassium hydrogen phosphate 2.5g, zinc sulfate 1mg, glucose 1g, salt
Sour pyridoxol 0.5mg, lecithin 1g, tetrachlorotetraiodofluorescein sodium 0.001g, glycerine 1ml, pH8.0, adds distilled water to 1000ml.
Preparation method：Each composition mixes in being formulated, and adjusts pH8.0, low pressure sterilizing, 4 DEG C of preservations.Main feature：Separate Pseudomonas aeruginosa life
It is long, do not allow or few other gram-Negative bacilluses grow, Pseudomonas aeruginosa is in green reaction, and green is obvious, easily observation.
The present invention has obvious as a kind of culture medium for promoting Pseudomonas aeruginosa fast breeding compared with conventional medium
Promote Pseudomonas aeruginosa growth, shorten the effect of incubation time.Shown with the medium culture Pseudomonas aeruginosa, the life of Pseudomonas aeruginosa logarithm
5-8 hours are advanced by for a long time, and stationary phase is grown.
Embodiment
The present invention is further described with reference to following instance.
Embodiment 1 prepares culture medium
(1) fast culture culture medium：By phenylacetic acid 1.2g, casein peptone 5g, magnesium sulfate 0.08g, sodium chloride 2g, phosphoric acid hydrogen
Dipotassium 2.5g, zinc sulfate 1mg, glucose 1g, puridoxine hydrochloride 0.5mg, lecithin 1g, tetrachlorotetraiodofluorescein sodium 0.001g,
Glycerine 1ml, pH8.0, add distilled water to 1000ml.Sterilizing, that is, obtain described culture medium.
(2) the conventional culture medium of prior art：
It is formulated (every liter)：Casein peptone 17g, soy peptone 3g, sodium chloride 5g, dipotassium hydrogen phosphate 2.5g, glucose
2.5g, lecithin 1g, Tween 80 7g, final pH 7.2 ± 0.2, sterilizing obtain described culture medium.
(3) common beef-protein medium.
The bacterium colony culture experiment of embodiment 2
By 2 × 106/ ml inoculum concentration is inoculated with Pseudomonas aeruginosa, Escherichia coli, salmonella, staphylococcus aureus respectively
In three kinds of culture mediums, put 37 degrees Celsius and cultivate 6 hours, determine bacterium number with haemocytometer counting method, experimental result is as follows：
| Bacterium is dense | Pseudomonas aeruginosa | Escherichia coli | Salmonella | Staphylococcus aureus |
| Culture medium 1 | 7.3×109/ml | 2.3×106/ml | 2.4×106/ml | 1.9×106/ml |
| Culture medium 2 | 4.5×108/ml | 5.2×109/ml | 4.1×109/ml | 1.0×109/ml |
| Culture medium 3 | 5.2×107/ml | 6.7×109/ml | 3.7×109/ml | 1.3×109/ml |
Data more than preferably promote Pseudomonas aeruginosa it can be shown that the fast culture media that the application provides can have
The characteristic of fast breeding, moreover, the culture medium is also not suitable for the breeding of other common bacteriums, be advantageous to the purifying of Pseudomonas aeruginosa
Culture.
The optimization of the medical sample Pseudomonas aeruginosa condition of culture of embodiment 3
Take medical sample secretion a little, be added in the test tube containing 3mL fast culture culture mediums, 42 DEG C first
220r/min cultivates 2h, and then 37 degrees Celsius of 220r/min culture 6h, nutrient solution color are changed into micro- green.Show in medical sample
Contain Pseudomonas aeruginosa.Also indicate that the bacterium in the nutrient solution is Pseudomonas aeruginosa by the conventional PCR identifications in this area.
By embodiment 3 it can be found that described cultural method relative to prior art the big rule of conventional cultural method
Mould shortens culture and the time differentiated, and a large amount of valuable times are saved for the inspection of germ.With applying valency well
Value.
Embodiment 4 and the obligate comparison of other Pseudomonas aeruginosa culture mediums
According to the method disclosed in CN1730649A, by CGMCC No.1445 bacterial strains with 2 × 106/ ml inoculum concentrations are inoculated into
In its dedicated liquid culture medium, the 220r ventilations culture 8h at 35 DEG C.
By CGMCC No.1445 bacterial strains with 2 × 106/ ml inoculum concentrations are inoculated into the fluid nutrient medium of the present invention, at 35 DEG C
Lower 220r ventilations culture 8h；As a result it is as follows：
| Bacterium is dense | Pseudomonas aeruginosa |
| Culture medium disclosed in CN1730649A | 2.1×109/ml |
| The application culture medium | 9.7×109/ml |
Result more than it can also be seen that the application culture medium in terms of the fast culture Pseudomonas aeruginosa on have it is preferable
Effect.
Claims (2)
- A kind of 1. culture medium, it is characterised in that：Energy fast culture Pseudomonas aeruginosa, is made up of following component：Phenylacetic acid 1.2g, junket egg White peptone 5g, magnesium sulfate 0.08g, sodium chloride 2g, dipotassium hydrogen phosphate 2.5g, zinc sulfate 1mg, glucose 1g, puridoxine hydrochloride 0.5mg, lecithin 1g, tetrachlorotetraiodofluorescein sodium 0.001g, glycerine 1ml, pH8.0, add distilled water to 1000ml.
- A kind of 2. method of fast culture Pseudomonas aeruginosa, it is characterised in that the medium culture green pus described in usage right requirement 1 Bacillus.
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| WO2007094000A2 (en) * | 2006-02-15 | 2007-08-23 | Botanocap Ltd. | Applications of microencapsulated essential oils |
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| CN103614451A (en) * | 2013-11-29 | 2014-03-05 | 中山鼎晟生物科技有限公司 | Method for detecting microorganisms of cosmetics |
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