CN103014130B - Staphylococcus xylosus selective culture medium and preparation method thereof - Google Patents
Staphylococcus xylosus selective culture medium and preparation method thereof Download PDFInfo
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- CN103014130B CN103014130B CN201310010767.8A CN201310010767A CN103014130B CN 103014130 B CN103014130 B CN 103014130B CN 201310010767 A CN201310010767 A CN 201310010767A CN 103014130 B CN103014130 B CN 103014130B
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Abstract
The invention discloses a staphylococcus xylosus selective culture medium and a preparation method thereof. The staphylococcus xylosus selective culture medium is characterized in that a formula for preparing 1000ml of the selective culture medium comprises the following components: 15g of malt extract, 150ml of egg protein fluid, 5g of sodium chloride, 0.5g of magnesium citrate, 15g of agar, 8-12mg of coetsoidin A, 0.4g of ambrettolide, 0.2g of monopotassium phosphate and distilled water added to 1000ml. The preparation method of the best optimization formula comprises the following steps of: dissolving the malt extract, the egg protein fluid, the sodium chloride, the magnesium citrate, the agar, the coetsoidin A, the ambrettolide, the monopotassium phosphate, L-cysteic acid and copper sulfate to Chinese yam water boiled liquid, uniformly mixing to be constant volume, and split charging after sterilization. Compared with the prior art, the method has the characteristics of being reliable to detect and high in separation rate of staphylococcus xylosus.
Description
Technical field
The present invention relates to field of medical examination, be specifically related to a kind of staphylococcus xylosus selective medium and preparation method thereof.
Background technology
Staphylococcus xylosus (Staphylococcus aureus) is a member in staphylococcus family, is low virulence conditioned pathogen, but recall rate and multi-drug resistant are and increase year by year trend in recent years, have become one of the main pathogenic fungi of hospital infection.Atypical symptom after staphylococcus xylosus infects, and present multidrug resistant phenomenon, bring certain difficulty to clinical diagnosis and treatment, the method for inspection of high efficient and reliable has been to need.
So-called microorganism culturing is a kind of technology that makes by artificial means microorganism growth breeding, and selectivity selective medium is for promoting or suppressing the organism (as cell or bacterium etc.) of certain type and the selective medium of design utilizes this selective medium needed microorganism can be separated from the microorganism mixing.
A on January 25th, 2012 is open, publication number is the substratum that the substratum of high-density culture of 102329747A < < staphylococcus xylosus A2 by name and the patent of invention of cultural method > > (application number is 201110258768.5) disclose a kind of high-density culture that is CGMCC No.5049 for staphylococcus xylosus (Staphylococcus xylosus) A2 culture presevation number, this formula comprises: 0.5 weight part lactose, 2 weight parts show peptone, 0.2 weight part yeast extract paste, 5 weight part mushroom juices, 7.5 weight part NaCl.This kind of substratum is for preparing starter for fermentative meat.But be applied to the effect that medical inspection aspect does not but reach cultivation, its reason is, medical inspection often contains other pathogenic bacterium with sample, especially golden Portugal bacterium, and this kind of substratum only provides the growth conditions of staphylococcus xylosus, but can not effectively remove the miscellaneous bacteria of sneaking into, cause the miscellaneous bacteria of sneaking into, as streptococcus aureus, faster than staphylococcus xylosus growth, covered very soon substratum, staphylococcus xylosus cannot grow.
Summary of the invention
Technical assignment of the present invention is for above the deficiencies in the prior art, and a kind of selective medium high to staphylococcus xylosus separation rate is provided.
The technical scheme that the present invention solves its technical problem is: a kind of staphylococcus xylosus selective medium, and its feature exists, and in the formula of preparation 1000ml selective medium, contains:
Fructus Hordei Germinatus soaks powder 15g;
Chicken Albumin liquid 150ml;
Sodium-chlor 5g;
Citrate of magnesia 0.5g;
Agar 15g;
Coetsoidin A 8 ~ 12mg;
Cyclohexa decen-7-olide 0.4 g;
Potassium primary phosphate 0.2g;
Distilled water adds to 1000ml.
In prioritization scheme, every 1000ml selective medium also contains copper sulfate 4mg.
In prioritization scheme, every 1000ml selective medium also contains Cys 0.1g.
In prioritization scheme, every 1000ml selective medium also contains Chinese yam water cooking liquid 200 ~ 300ml of concentration 200g/L.
In optimization scheme, every 1000ml selective medium also contains copper sulfate 4mg; Cys 0.1g; Chinese yam water cooking liquid 200 ~ 300ml of concentration 200g/L.
A preparation method for staphylococcus xylosus selective medium, is characterized in that comprising the following steps:
(1) get Chinese yam and add water boil 1 hour, filter, get filtrate, adjusting volume is 200g/L to concentration, obtains Chinese yam water cooking liquid;
(2) get Fructus Hordei Germinatus and soak powder; Chicken Albumin liquid; Sodium-chlor; Citrate of magnesia; Agar; Coetsoidin A; Cyclohexa decen-7-olide; Potassium primary phosphate; Cys and copper sulfate are dissolved in step 1 gained Chinese yam water cooking liquid, mix after filtration distilled water constant volume, temperature sterilization 2 times, each 45 min, packing after sterilizing.
Wherein said Fructus Hordei Germinatus soaks powder, Chicken Albumin liquid provides carbon nitrogen source and energy; Citrate of magnesia and potassium primary phosphate are buffer reagent, and can suppress varied bacteria growing; Cyclohexa decen-7-olide, Cys belong to amino acids, are nutrition agent; And cyclohexa decen-7-olide is staphylococcus xylosus specificity nutrition agent, can specific promotion staphylococcus xylosus breeding.Agar is the peptizer of selective medium; Coetsoidin A has special bacteriostatic action for golden Portugal bacterium, invalid for staphylococcus xylosus; Chicken Albumin liquid contains each peptide species, fat, somatomedin, hormone etc., and these materials are to promoting that the Growth and Reproduction of staphylococcus xylosus is all very important factor, and Chicken Albumin liquid also has the constant effect of selective medium pH value that maintains in addition.Cupric ion belongs to heavy metal ion, under this concentration dose of 4mg/L, be deposited on golden Portugal bacterium inner, cause cell wall to disappear or attenuation, rrna is gathered into agglomerate, finally causes bacterium distortion and breaks, present the property damage of heavy metal source, and staphylococcus xylosus to the transportcapacity of heavy metal and penetrating power lower than golden Portugal bacterium, be also that the resistance of heavy metal is higher, so add copper sulfate can effectively suppress golden Portugal bacteria growing.Chinese yam is the dry rhizome of Dioscoreaceae plant Chinese yam Dioscorea opposita Thunb., contain the Multiple components such as protein, fat, starch, also first sugar, amino acid, energy metabolism, cell development, the metabolic factor can be provided, especially Chinese yam stem tuber is containing diosgenin (diosgenin) and polyphenoloxidase, help lend some impetus to road Deng's aureus growth, the especially growth of bacterium colony under heavy metal environment.
The present invention compared with prior art, has the feature reliable, staphylococcus xylosus separation rate is high that detects.
Embodiment
Below in conjunction with practical situation, the specific embodiment of the present invention is elaborated.
Embodiment 1, contains: Fructus Hordei Germinatus soaks powder 15g in the formula of preparation 1000ml selective medium; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 8mg; Cyclohexa decen-7-olide 0.4 g; Potassium primary phosphate
0.2g; Distilled water adds to 1000ml.Preparation method: get Fructus Hordei Germinatus and soak powder; Chicken Albumin liquid; Sodium-chlor; Citrate of magnesia; Agar; Coetsoidin A; Cyclohexa decen-7-olide; Potassium primary phosphate is dissolved in distilled water, mixes after filtration distilled water constant volume, temperature sterilization 2 times, each 45 min, packing after sterilizing.
Embodiment 2, contain: Fructus Hordei Germinatus soaks powder 15g in the formula of preparation 1000ml selective medium; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 12mg; Cyclohexa decen-7-olide 0.4 g; Potassium primary phosphate
0.2g; Copper sulfate 4mg; Distilled water adds to 1000ml.Preparation method: get Fructus Hordei Germinatus and soak powder; Chicken Albumin liquid; Sodium-chlor; Citrate of magnesia; Agar; Coetsoidin A; Cyclohexa decen-7-olide; Potassium primary phosphate; Copper sulfate is dissolved in distilled water, mixes after filtration distilled water constant volume, temperature sterilization 2 times, each 45 min, packing after sterilizing.
Embodiment 3, contain: Fructus Hordei Germinatus soaks powder 15g in the formula of preparation 1000ml selective medium; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 10mg; Cyclohexa decen-7-olide 0.4 g; Potassium primary phosphate
0.2g; Cys
0.1g; Distilled water adds to 1000ml.Preparation method: get Fructus Hordei Germinatus and soak powder; Chicken Albumin liquid; Sodium-chlor; Citrate of magnesia; Agar; Coetsoidin A; Cyclohexa decen-7-olide; Potassium primary phosphate; Cys is dissolved in distilled water, mixes after filtration distilled water constant volume, temperature sterilization 2 times, each 45 min, packing after sterilizing.
Embodiment 4, contain: Fructus Hordei Germinatus soaks powder 15g in the formula of preparation 1000ml selective medium; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 9mg; Cyclohexa decen-7-olide 0.4 g; Potassium primary phosphate
0.2g; The Chinese yam water cooking liquid 200ml of concentration 200g/L; Distilled water adds to 1000ml.Preparation method:
(1) get Chinese yam and add water boil 1 hour, filter, get filtrate, adjusting volume is 200g/L to concentration, obtains Chinese yam water cooking liquid;
(2) get Fructus Hordei Germinatus and soak powder; Chicken Albumin liquid; Sodium-chlor; Citrate of magnesia; Agar; Coetsoidin A; Cyclohexa decen-7-olide; Potassium primary phosphate; Be dissolved in step 1 gained Chinese yam water cooking liquid, mix after filtration, distilled water constant volume, temperature sterilization 2 times, each 45 min, packing after sterilizing.
Embodiment 5, contain: Fructus Hordei Germinatus soaks powder 15g in the formula of preparation 1000ml selective medium; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 11mg; Cyclohexa decen-7-olide 0.4 g; Potassium primary phosphate
0.2g; Copper sulfate 4mg; Cys
0.1g; The Chinese yam water cooking liquid 300ml of concentration 200g/L; Distilled water adds to 1000ml.Preparation method is:
(1) get Chinese yam and add water boil 1 hour, filter, get filtrate, adjusting volume is 200g/L to concentration, obtains Chinese yam water cooking liquid;
(2) get Fructus Hordei Germinatus and soak powder; Chicken Albumin liquid; Sodium-chlor; Citrate of magnesia; Agar; Coetsoidin A; Cyclohexa decen-7-olide; Potassium primary phosphate; Cys and copper sulfate are dissolved in step 1 gained Chinese yam water cooking liquid, mix after filtration distilled water constant volume, temperature sterilization 2 times, each 45 min, packing after sterilizing.
Described Chinese yam water cooking liquid concentration is 200g/L, refers in every liter of Chinese yam water cooking liquid and contains crude drug 200g.
Gained staphylococcus xylosus selective medium of the present invention has and detects the feature reliable, staphylococcus xylosus separation rate is high, and for clinical data fully proves, pertinent data is as follows.
1 object and method.
1.1 selective medium preparations: establish altogether 3 groups, be respectively control group, embodiment 1 scheme group, embodiment 5 scheme groups.Control group is staphylococcus xylosus A2 high-density culture medium, and filling a prescription is: 0.5g lactose, 2g show that peptone, 0.2g yeast extract paste, 5g mushroom juice, 7.5gNaCl are dissolved in 1000g distilled water.Formula and the preparation method of embodiment 1 scheme group, embodiment 5 scheme groups are shown in embodiment.
1.2 methods: by sneaking into gold-coloured staphylococci in 50 parts of samples of determining the staphylococcus xylosus positive via PCR, be inoculated on substratum, hatch at 37 ℃ 24 hours.
1.3 statistical analysis: carry out statistical study with SPSS 13.0, P< 0.05 indicates significant.
2 results: in 50 parts of inoculation samples, microbial culture is separated and be accredited as staphylococcus xylosus, and embodiment 1 scheme group is 48 examples, and embodiment 5 scheme groups are 49 examples.Control group staphylococcus xylosus A2 high-density culture medium, through being accredited as totally 23 examples of staphylococcus xylosus, being learned and is processed by statistics, and various embodiments of the present invention group and control group relatively have notable difference (P<0.05).Result shows, embodiment of the present invention staphylococcus xylosus separation rate is apparently higher than existing selective medium.
Claims (1)
1. a staphylococcus xylosus selective medium, its feature exists, and contains: Fructus Hordei Germinatus soaks powder 15g in the formula of preparation 1000ml selective medium; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 11mg; Cyclohexa decen-7-olide 0.4 g; Potassium primary phosphate 0.2g; Copper sulfate 4mg; Cys 0.1g; The Chinese yam water cooking liquid 300ml of concentration 200g/L; Distilled water adds to 1000ml.
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Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1297655C (en) * | 2003-06-26 | 2007-01-31 | 河南双汇投资发展股份有限公司 | Staphylococcus xylosus I2 strain, composite ferment produced thereby and the use of ferment in meat ware |
| JP4931351B2 (en) * | 2005-01-12 | 2012-05-16 | 有限会社 ミクロデント | 3D fixed medium |
| CN101469340B (en) * | 2007-12-26 | 2012-05-02 | 上海复星佰珞生物技术有限公司 | Identification plate for gram positive aerobic bacteria and preparation thereof |
| CN102329747B (en) * | 2011-09-02 | 2012-12-26 | 东北农业大学 | Culture medium and culture method for high-density culture of Staphylococcus xylosus A2 |
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Effective date of registration: 20140115 Address after: 262100 Anqiu Municipal People's Hospital, Anqiu health Road, Shandong, 246, China Applicant after: Zhang Yueqiang Address before: 276826 Rizhao City traditional Chinese medicine hospital, 35 Hai Lu, Shandong, Rizhao City Applicant before: Hou Sujun |
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