CN103014127B - Staphylococcus lentus selective medium and preparation method thereof - Google Patents
Staphylococcus lentus selective medium and preparation method thereof Download PDFInfo
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- CN103014127B CN103014127B CN2013100107502A CN201310010750A CN103014127B CN 103014127 B CN103014127 B CN 103014127B CN 2013100107502 A CN2013100107502 A CN 2013100107502A CN 201310010750 A CN201310010750 A CN 201310010750A CN 103014127 B CN103014127 B CN 103014127B
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Abstract
The invention discloses a staphylococcus lentus selective medium and a preparation method thereof. The staphylococcus lentus selective medium is characterized in that the formula of 1000 ml of selective medium comprises the following components by weight: 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, 3.6g of N-acetyl-D-glucosamine and 0.2g of monopotassium phosphate, which are prepared to 1000 ml by adding distilled water. The optimal preparation method for the staphylococcus lentus selective medium comprises the following steps of: dissolving the malt extract, the egg protein fluid, sodium chloride, magnesium citrate, agar, coetsoidin, N-acetyl-D-glucosamine, monopotassium phosphate, L-cysteine and copper sulfate in lily decocted liquid; uniformly mixing the components, metering the volume of the mixture, sterilizing, splitting and packaging the mixture. Compared with the prior art, the preparation method has the characteristics that the detection is reliable, and the separation rate of the staphylococcus lentus is high.
Description
Technical field
The present invention relates to field of medical examination, be specifically related to a kind of slow staphylococcus selective medium and preparation method thereof.
Background technology
Slowly staphylococcus (Staphylococcus lentus) belongs to Staphylococcus (Staphylococcus).This bacterium is independently planted its of being divided into Staphylococcus in nineteen eighty-three by Schleifer K H etc., i.e. slow staphylococcus (S.lentus).Slowly staphylococcus once was considered to not have pathogenic, but increasing data shows that S.lentus is the perhaps very important pathogenic bacterium of many animals of the mankind, can cause human diseases and occupy critical role in hospital infection, especially for newborn child, the crowd that has serious underlying diseases or accept for a long time the hypoimmunities such as patient of hospitalize, more easily cause serious consequence.Because the multidrug resistant problem of S.lentus is day by day serious, the infection that S.lentus causes is the trend of remarkable increase at home and abroad.
So-called microorganism culturing is a kind of technology that makes by artificial means the microorganism growth breeding, and the selectivity selective medium is the organism (as cell or bacterium etc.) that promotes or suppress certain type and the selective medium of design utilizes this selective medium needed microorganism can be separated from the microorganism that mixes.
The staphylococcus selective medium that generally uses clinically at present is manitol salt agar (Manitol Salt Agar), and the selective separation that is used for streptococcus aureus is cultivated.Medium component is: peptone, Chicken Albumin liquid, PEARLITOL 25C, sodium-chlor, phenol red, agar.The streptococcus aureus bacterium colony is yellow, and intestinal bacteria and Salmonellas bacterium colony are colourless.But this kind substratum can not selectivity be cultivated slow staphylococcus, and its reason is that streptococcus aureus is faster than slow aureus growth in present selective medium, has covered very soon substratum, and slowly staphylococcus can't grow.
Summary of the invention
Technical assignment of the present invention is for above the deficiencies in the prior art, provides a kind of to the high selective medium of slow staphylococcus separation rate.
The technical scheme that the present invention solves its technical problem is: a kind of slow staphylococcus selective medium, and its feature exists, and contain in the formula of preparation 1000ml selective medium: Fructus Hordei Germinatus soaks powder 15g; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 8 ~ 12mg; N-ethanoyl-D-Glucose amine 3.6 g; Potassium primary phosphate 0.2g; Distilled water adds to 1000ml.
In prioritization scheme, every 1000ml selective medium also contains copper sulfate 5mg.
In prioritization scheme, every 1000ml selective medium also contains Cys 0.1g.
In prioritization scheme, every 1000ml selective medium also contains lily water cooking liquid 200 ~ 300ml of concentration 200g/L.
In optimization scheme, every 1000ml selective medium also contains copper sulfate 5mg; Cys 0.1g; Lily water cooking liquid 200 ~ 300ml of concentration 200g/L.
A kind of preparation method of slow staphylococcus selective medium is characterized in that comprising the following steps:
(1) get lily and added water boil 1 hour, filter, get filtrate, adjusting volume is 200g/L to concentration, obtains the lily water cooking liquid;
(2) get Fructus Hordei Germinatus and soak powder; Chicken Albumin liquid; Sodium-chlor; Citrate of magnesia; Agar; Coetsoidin A; N-ethanoyl-D-Glucose amine; Potassium primary phosphate; Cys and copper sulfate are dissolved in step 1 gained lily water cooking liquid, after mixing filtration, and distilled water constant volume, temperature sterilization 2 times, each 45 min, sterilizing rear packing.
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; Cys belongs to amino acids, is nutrition agent; Agar is the peptizer of selective medium; Coetsoidin A has special bacteriostatic action for golden Portugal bacterium, and is invalid for slow staphylococcus; N-ethanoyl-D-Glucose amine is slow aureus specific nutrition agent, can the slow staphylococcus breeding of specific promotion.Chicken Albumin liquid contains each peptide species, fat, somatomedin, hormone etc., and these materials are to promoting that slow staphylococcic Growth and Reproduction is all very important factor, and Chicken Albumin liquid also has the constant effect of selective medium pH value of keeping in addition.Cupric ion belongs to heavy metal ion, under this concentration dose of 5mg/L, be deposited on golden Portugal bacterium inner, cause cell wall to disappear or attenuation, rrna is gathered into agglomerate, finally causes the bacterium distortion and breaks, present the property damage of heavy metal source, and slowly staphylococcus 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.Lily is the dry meat scale leaf of liliaceous plant tiger lily Lilium lancifolium Thunb., lily Lilium brownii F.E.Brown var. viridulum Baker or Lilium tenuifolium Lilium pumilum DC., contain the Multiple components such as protein, fat, starch, also first sugar, VITMAIN B1, B2, pantothenic acid, vitamins C, β-carotene, energy metabolism, cell development, the metabolic factor can be provided, help lend some impetus to slow aureus growth, the especially growth of bacterium colony under the heavy metal environment.
The present invention compared with prior art, has reliable, the slow high characteristics of staphylococcus separation rate that detect.
Embodiment
Below in conjunction with practical situation, the specific embodiment of the present invention is elaborated.
Embodiment 1, and contain in the formula of preparation 1000ml selective medium: Fructus Hordei Germinatus soaks powder 15g; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 8mg; N-ethanoyl-D-Glucose amine 3.6 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; N-ethanoyl-D-Glucose amine; Potassium primary phosphate is dissolved in distilled water, after mixing filtration, and distilled water constant volume, temperature sterilization 2 times, each 45 min, sterilizing rear packing.
Embodiment 2, and contain in the formula of preparation 1000ml selective medium: Fructus Hordei Germinatus soaks powder 15g; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 12mg; N-ethanoyl-D-Glucose amine 3.6 g; Potassium primary phosphate
0.2g； Copper sulfate 5mg; 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; N-ethanoyl-D-Glucose amine; Potassium primary phosphate; Copper sulfate is dissolved in distilled water, after mixing filtration, and distilled water constant volume, temperature sterilization 2 times, each 45 min, sterilizing rear packing.
Embodiment 3, and contain in the formula of preparation 1000ml selective medium: Fructus Hordei Germinatus soaks powder 15g; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 10mg; N-ethanoyl-D-Glucose amine 3.6 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; N-ethanoyl-D-Glucose amine; Potassium primary phosphate; Cys is dissolved in distilled water, after mixing filtration, and distilled water constant volume, temperature sterilization 2 times, each 45 min, sterilizing rear packing.
Embodiment 4, and contain in the formula of preparation 1000ml selective medium: Fructus Hordei Germinatus soaks powder 15g; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 9mg; N-ethanoyl-D-Glucose amine 3.6 g; Potassium primary phosphate
0.2g； The lily water cooking liquid 200ml of concentration 200g/L; Distilled water adds to 1000ml.The preparation method:
(1) get lily and added water boil 1 hour, filter, get filtrate, adjusting volume is 200g/L to concentration, obtains the lily water cooking liquid;
(2) get Fructus Hordei Germinatus and soak powder; Chicken Albumin liquid; Sodium-chlor; Citrate of magnesia; Agar; Coetsoidin A; N-ethanoyl-D-Glucose amine; Potassium primary phosphate; Be dissolved in step 1 gained lily water cooking liquid, after mixing filtration, distilled water constant volume, temperature sterilization 2 times, each 45 min, sterilizing rear packing.
Embodiment 5, and contain in the formula of preparation 1000ml selective medium: Fructus Hordei Germinatus soaks powder 15g; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 11mg; N-ethanoyl-D-Glucose amine 3.6 g; Potassium primary phosphate
0.2g； Copper sulfate 5mg; Cys
0.1g； The lily water cooking liquid 300ml of concentration 200g/L; Distilled water adds to 1000ml.The preparation method is:
(1) get lily and added water boil 1 hour, filter, get filtrate, adjusting volume is 200g/L to concentration, obtains the lily water cooking liquid;
(2) get Fructus Hordei Germinatus and soak powder; Chicken Albumin liquid; Sodium-chlor; Citrate of magnesia; Agar; Coetsoidin A; N-ethanoyl-D-Glucose amine; Potassium primary phosphate; Cys and copper sulfate are dissolved in step 1 gained lily water cooking liquid, after mixing filtration, and distilled water constant volume, temperature sterilization 2 times, each 45 min, sterilizing rear packing.
Described lily water cooking liquid concentration is 200g/L, refers in every liter of lily water cooking liquid and contains crude drug 200g.
The slow staphylococcus selective medium of gained of the present invention has and detects reliable, slow high characteristics of staphylococcus separation rate, is the clinical data sufficient proof, and pertinent data is as follows.
1 object and method.
1.1 selective medium preparation: establish altogether 3 groups, be respectively control group, embodiment 1 scheme group, embodiment 5 scheme groups.Control group is staphylococcus selective agar 110(CHAPMAN agar), filling a prescription is: pancreas casein peptone, yeast soak powder, gelatin, PEARLITOL 25C, lactose, sodium-chlor, dipotassium hydrogen phosphate, agar.Formula and the preparation method of embodiment 1 scheme group, embodiment 5 scheme groups see embodiment.
1.2 method:, with sneaking into gold-coloured staphylococci in 30 parts of samples of determining the slow staphylococcus positive via PCR, be inoculated on substratum, hatched under 37 ℃ 24 hours.
1.3 statistical analysis: carry out statistical study with SPSS 13.0, P＜0.05 expression has significant.
2 results: in 30 parts of inoculation samples, microbial culture is separated and is accredited as slowly staphylococcicly, and embodiment 1 scheme group is 25 examples, and embodiment 5 scheme groups are 26 examples.Control group CHAPMAN nutrient agar bacterium colony is golden Portugal bacterium through identifying to be proved finally, learns and processes by statistics, and various embodiments of the present invention group and control group relatively have notable difference (P＜0.001).Result shows, the slow staphylococcus separation rate of the embodiment of the present invention is apparently higher than existing selective medium.
Claims (2)
1. slow staphylococcus selective medium, its feature exists, and contain in the formula of preparation 1000ml selective medium: Fructus Hordei Germinatus soaks powder 15g; Chicken Albumin liquid 150ml; Sodium-chlor 5g; Citrate of magnesia 0.5g; Agar 15g; Coetsoidin A 8 ~ 12mg; N-ethanoyl-D-Glucose amine 3.6 g; Potassium primary phosphate 0.2g; Distilled water adds to 1000ml.
2. a kind of slow staphylococcus selective medium according to claim 1, its feature exists: every 1000ml selective medium also contains copper sulfate 5mg; Cys 0.1g; Lily water cooking liquid 200 ~ 300ml of concentration 200g/L.
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Citations (2)
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| CN102851230A (en) * | 2012-04-18 | 2013-01-02 | 湖北中烟工业有限责任公司 | Preparation method and application of Bacillus methylotrophicus and tobacco chaff extract thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102851230A (en) * | 2012-04-18 | 2013-01-02 | 湖北中烟工业有限责任公司 | Preparation method and application of Bacillus methylotrophicus and tobacco chaff extract thereof |
| CN102747008A (en) * | 2012-07-23 | 2012-10-24 | 湖北中烟工业有限责任公司 | Bacillus methylotrophicus VJ4-1 and method for producing natural vanillin by ferulic acid biotransformation with the same |
Non-Patent Citations (4)
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| 仓鼠源缓慢葡萄球菌的分离鉴定及生物学特性研究;盛相鹏等;《动物医学进展》;20101231;第31卷(第7期);第30-35页 * |
| 王沛等.荧光原位杂交法快速鉴定金黄色葡萄球菌和.《中华检验医学杂志》.2004,第27卷(第7期),第434-436页. |
| 盛相鹏等.仓鼠源缓慢葡萄球菌的分离鉴定及生物学特性研究.《动物医学进展》.2010,第31卷(第7期),第30-35页. |
| 荧光原位杂交法快速鉴定金黄色葡萄球菌和;王沛等;《中华检验医学杂志》;20040731;第27卷(第7期);第434-436页 * |
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Effective date of registration: 20131014 Address after: 276826 Rizhao People's Hospital, 126 Tai'an Road, Shandong, Rizhao City, China Applicant after: Dong Yanjun Address before: 276826 Rizhao City traditional Chinese medicine hospital, 35 Hai Lu, Shandong, Rizhao City Applicant before: Hou Sujun |
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