CN101659980B - Selective coloration culture medium of clostridium perfringens - Google Patents
Selective coloration culture medium of clostridium perfringens Download PDFInfo
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- CN101659980B CN101659980B CN2009101677827A CN200910167782A CN101659980B CN 101659980 B CN101659980 B CN 101659980B CN 2009101677827 A CN2009101677827 A CN 2009101677827A CN 200910167782 A CN200910167782 A CN 200910167782A CN 101659980 B CN101659980 B CN 101659980B
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Abstract
The invention relates to a selective coloration culture medium for fast culturing and identifying clostridium perfringens. The selective coloration culture medium is characterized in that the preparation of 1000ml of the culture medium requires 12.0g-20.0g of casease hydrolysate, 6.0g-12.0g of yeast extract powder, 0.20g-0.80g of sodium sulfite, 0.005g-0.015g of polymyxin B, 0.05g-0.20g of sulfadiazine, 0.20g-0.80g of ferric citrate, 4-12 units of Y.S.N. and 10.0g-18.0g of agar powder and also requires the final addition of distilled water till the volume of 1000ml is achieved. The selective coloration culture medium has the advantages that the culturing time is obviously reduced when being compared with a conventional method and other bacteria with the same culturing characteristics are inhibited and do not grow so that the effect of fast clostridium perfringens culturing and identification can be realized.
Description
Technical field:
The present invention relates to a kind of fast culture and selective coloration culture medium of identifying clostridium perfringens of being used for.
Background technology:
Clostridium perfringens is distributed widely in the enteron aisle of nature and humans and animals, and can cause the multiple disease of humans and animals, and modal clinically is gas gangrene and the food poisoning that is caused by clostridium perfringens.Gas gangrene is that pathogenic bacteria is invaded the serious acute infection that wound causes, poisoning with tissue necrosis, oedema, flatulence, whole body is characteristic, good sending out in lower limb, and mortality ratio is up to 40%-100%.This disease often is the polyinfection that two or more bacteriums are participated in, and wherein 60%-80% is that clostridium perfringens causes.Therefore, in early days apace from sample separation and Culture go out this bacterium, seem very important for the diagnosis and the treatment of disease.Clostridium perfringens is the thick bacillus of Gram-positive, and anaerobism but less demanding also can be grown in the environment of a small amount of aerobic is arranged, and nutritional requirement is not high, and growth and breeding is rapid.Traditional culture identification method; Be with specimen inoculation on blood agar plate and egg yolk agar flat board; Under 37 ℃ of conditions, cultivated 18-24 hour; Select suspicious bacterium colony and continue to inoculate differential medium continuation cultivation again, or increase the bacterium cultivation after 8-10 hour, change kind of blood agar plate and egg yolk agar flat board at cooked meat medium.The traditional method length that expends time in, other bacterium similar with the clostridium perfringens cultural characters also can grow, and makes the differential diagnosis lack of specific, and making the preliminary evaluation diagnosis often needs more than 24 hours.
Summary of the invention:
In view of above present situation, the present invention provides a kind of selective coloration culture medium of clostridium perfringens, and the incubation time that has solved above-mentioned traditional culture identification method existence is long, lacks the problem of specificity.
Technical scheme of the present invention realizes through following content:
A kind of selective coloration culture medium of clostridium perfringens is characterized in that: the composition of configuration 1000ml substratum is rennase hydrolyzate 12.0g-20.0g, and yeast soaks powder 6.0g-12.0g; S-WAT 0.20g-0.80g, PXB 0.005g-0.015g, Sulphadiazine Sodium 0.05g-0.20g; Ironic citrate 0.20g-0.80g; 4-12 unit of pancreas Suning, agar powder 10.0g-18.0g, adding distil water is to 1000ml.
The composition of said configuration 1000ml substratum is rennase hydrolyzate 12.0g-14.0g, and yeast soaks powder 10.0g-12.0g, S-WAT 0.20g-0.40g; PXB 0.010g-0.015g; Sulphadiazine Sodium 0.05g-0.10g, ironic citrate 0.60g-0.80g, 4-6 unit of pancreas Suning; Agar powder 16.0g-18.0g, adding distil water is to 1000ml.
The composition of said configuration 1000ml substratum is rennase hydrolyzate 14.0g-16.0g, and yeast soaks powder 8.0g-10.0g, S-WAT 0.40g-0.60g; PXB 0.008g-0.010g; Sulphadiazine Sodium 0.10g-0.15g, ironic citrate 0.40g-0.60g, 6-10 unit of pancreas Suning; Agar powder 12.0g-16.0g, adding distil water is to 1000ml.
The composition of said configuration 1000ml substratum is rennase hydrolyzate 16.0g-20.0g, and yeast soaks powder 6.0g-8.0g, S-WAT 0.60g-0.80g; PXB 0.005g-0.008g; Sulphadiazine Sodium 0.15g-0.20g, ironic citrate 0.20g-0.40g, 10-12 unit of pancreas Suning; Agar powder 10.0g-12.0g, adding distil water is to 1000ml.
Said zero(ppm) water is distilled water.
Said substratum is through high pressure steam sterilization, and its force value is 103.4kPa.
The final pH value of said substratum is 7.0 ± 0.2 under 25 ℃ of conditions.
Layoutprocedure: under 35-37 ℃ temperature condition; Add an amount of distilled water earlier to container; After again rennase hydrolyzate, yeast being soaked powder, S-WAT, PXB, Sulphadiazine Sodium, ironic citrate, pancreas Suning, agar powder and add in the distilled water fully dissolving; Be settled to 1000ml with volumetric flask, making its final pH value under 25 ℃ through interpolation acid (HCL) or alkaline matter (NaOH) in solution is 7.0 ± 0.2, at last through 103.4kPa (1.05kg/cm
3) high pressure steam sterilization, need be during packing, and be sub-packed in aseptic plate at Bechtop.
Advantage of the present invention: incubation time obviously shortens than traditional method, and other bacterium with identical cultural characters is suppressed all and does not grow, thereby has realized fast culture and the effect of identifying clostridium perfringens.
Embodiment:
Under 35-37 ℃ of condition; Add an amount of distilled water earlier to container; After again rennase hydrolyzate, yeast being soaked powder, S-WAT, PXB, Sulphadiazine Sodium, ironic citrate, pancreas Suning, agar powder and add in the distilled water fully dissolving; Be settled to 1000ml with volumetric flask, making its final pH value under 25 ℃ through the acid perhaps alkaline matter of interpolation in solution is 7.0 ± 0.2, at last through 103.4kPa (1.05kg/cm
3) high pressure steam sterilization, need be during packing, and be sub-packed in aseptic plate at Bechtop.
The rennase hydrolyzate, yeast soaks powder, and the required nitrogenous source of bacterial growth, carbon source and growth factor are provided; Agar powder is the vehicle of substratum; PXB, Sulphadiazine Sodium suppresses other varied bacteria growing; Pancreas Suning is a positive growth factor; Ironic citrate and S-WAT are colored indicator, and S-WAT also can suppress other Gram-positive bacteria growing.
Cultivated 8-12 hour at 35-37 ℃ with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.
Embodiment 1
A kind of substratum is provided, is used for fast culture and identifies clostridium perfringens.Staple is: rennase hydrolyzate 12.0g, yeast soaks powder 12.0g, S-WAT 0.20g, PXB 0.015g, Sulphadiazine Sodium 0.05g, ironic citrate 0.80g, 4 units of pancreas Suning, agar powder 18.0g.The accurate above composition of weighing after fully dissolving with distilled water, is settled to 1000ml with volumetric flask, at 103.4kPa (1.05kg/cm
3) condition under carry out high pressure steam sterilization, it is subsequent use to be sub-packed in aseptic plate, final PH (25 ℃) value is 6.8.Cultivated 8 hours under 35 ℃ of conditions with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.
Embodiment 2
A kind of substratum is provided, is used for fast culture and identifies clostridium perfringens.Staple is: rennase hydrolyzate 20.0g, yeast soaks powder 6.0g, S-WAT 0.80g, PXB 0.005g, Sulphadiazine Sodium 0.20g, ironic citrate 0.20g, 12 units of pancreas Suning, agar powder 10.0g.The accurate above composition of weighing after fully dissolving with distilled water, is settled to 1000ml with volumetric flask, at 103.4kPa (1.05kg/cm
3) condition under carry out high pressure steam sterilization, it is subsequent use to be sub-packed in aseptic plate, final PH (25 ℃) value is 7.2.Cultivated 12 hours under 37 ℃ of conditions with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.
Embodiment 3
A kind of substratum is provided, is used for fast culture and identifies clostridium perfringens.Staple is: rennase hydrolyzate 14.0g, yeast soaks powder 10.0g, S-WAT 0.40g, PXB 0.01g, Sulphadiazine Sodium 0.10g, ironic citrate 0.60g, 6 units of pancreas Suning, agar powder 16.0g.The accurate above composition of weighing after fully dissolving with distilled water, is settled to 1000ml with volumetric flask, at 103.4kPa (1.05kg/cm
3) condition under carry out high pressure steam sterilization, it is subsequent use to be sub-packed in aseptic plate, final PH (25 ℃) value is 7.0.Cultivated 10 hours under 36 ℃ of conditions with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.
Embodiment 4
A kind of substratum is provided, is used for fast culture and identifies clostridium perfringens.Staple is: rennase hydrolyzate 16.0g, yeast soaks powder 8.0g, S-WAT 0.60g, PXB 0.008g, Sulphadiazine Sodium 0.10g, ironic citrate 0.40g, 10 units of pancreas Suning, agar powder 12.0g.The accurate above composition of weighing after fully dissolving with distilled water, is settled to 1000ml with volumetric flask, at 103.4kPa (1.05kg/cm
3) condition under carry out high pressure steam sterilization, it is subsequent use to be sub-packed in aseptic plate, final PH (25 ℃) value is 6.9.Cultivated 9 hours under 36 ℃ of conditions with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.
Embodiment 5
A kind of substratum is provided, is used for fast culture and identifies clostridium perfringens.Staple is: rennase hydrolyzate 18.0g, yeast soaks powder 10.0g, S-WAT 0.60g, PXB 0.012g, Sulphadiazine Sodium 0.15g, ironic citrate 0.60g, 10 units of pancreas Suning, agar powder 16.0g.The accurate above composition of weighing after fully dissolving with distilled water, is settled to 1000ml with volumetric flask, at 103.4kPa (1.05kg/cm
3) condition under carry out high pressure steam sterilization, it is subsequent use to be sub-packed in aseptic plate, final PH (25 ℃) value is 7.1.Cultivated 11 hours under 37 ℃ of conditions with this substratum, the clostridium perfringens well-grown, bacterium colony shows black, and the Gram-positive and the negative bacterium of other reduction sulphite are not grown, thereby realize fast culture and identify clostridium perfringens.
Claims (7)
1. selective coloration culture medium of clostridium perfringens is characterized in that: the composition of configuration 1000ml substratum is rennase hydrolyzate 12.0g-20.0g, and yeast soaks powder 6.0g-12.0g; S-WAT 0.20g-0.80g, PXB 0.005g-0.015g, Sulphadiazine Sodium 0.05g-0.20g; Ironic citrate 0.20g-0.80g; 4-12 unit of pancreas Suning, agar powder 10.0g-18.0g, adding distil water is to 1000ml.
2. selective coloration culture medium of clostridium perfringens according to claim 1 is characterized in that: the composition of configuration 1000ml substratum is rennase hydrolyzate 12.0g-14.0g, and yeast soaks powder 10.0g-12.0g; S-WAT 0.20g-0.40g, PXB 0.010g-0.015g, Sulphadiazine Sodium 0.05g-0.10g; Ironic citrate 0.60g-0.80g; 4-6 unit of pancreas Suning, agar powder 16.0g-18.0g, adding distil water is to 1000ml.
3. selective coloration culture medium of clostridium perfringens according to claim 1 is characterized in that: the composition of configuration 1000ml substratum is rennase hydrolyzate 14.0g-16.0g, and yeast soaks powder 8.0g-10.0g; S-WAT 0.40g-0.60g, PXB 0.008g-0.010g, Sulphadiazine Sodium 0.10g-0.15g; Ironic citrate 0.40g-0.60g; 6-10 unit of pancreas Suning, agar powder 12.0g-16.0g, adding distil water is to 1000ml.
4. selective coloration culture medium of clostridium perfringens according to claim 1 is characterized in that: the composition of configuration 1000ml substratum is rennase hydrolyzate 16.0g-20.0g, and yeast soaks powder 6.0g-8.0g; S-WAT 0.60g-0.80g, PXB 0.005g-0.008g, Sulphadiazine Sodium 0.15g-0.20g; Ironic citrate 0.20g-0.40g; 10-12 unit of pancreas Suning, agar powder 10.0g-12.0g, adding distil water is to 1000ml.
5. according to any described selective coloration culture medium of clostridium perfringens among the claim 1-4, it is characterized in that: said zero(ppm) water is distilled water.
6. according to any described selective coloration culture medium of clostridium perfringens among the claim 1-4, it is characterized in that: said substratum is through high pressure steam sterilization, and its force value is 103.4kPa.
7. according to any described selective coloration culture medium of clostridium perfringens among the claim 1-4, it is characterized in that: the final pH value of said substratum is 7.0 ± 0.2 under 25 ℃ of conditions.
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| CN103343158B (en) * | 2013-07-20 | 2014-05-14 | 吉林农业大学 | Detection paper piece for clostridium perfringens, preparation method and application for same |
| CN103966142B (en) * | 2014-05-21 | 2016-05-18 | 福建省农业科学院 | A kind of culture medium and preparation method who reduces Pasteurella multocida pod membrane production cost |
| CN104178551A (en) * | 2014-09-09 | 2014-12-03 | 青岛润鑫伟业科贸有限公司 | Reinforced clostridium identification agar culture medium and application thereof |
| CN108102981B (en) * | 2018-02-07 | 2020-08-04 | 江南大学 | Culture medium for improving clostridium butyricum screening efficiency |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| SU1751987A1 (en) * | 1990-02-07 | 1996-01-10 | Ленинградский научно-исследовательский институт травматологии и ортопедии им.Р.Р.Вредена | Nutrient medium for clostridia isolation |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| SU1751987A1 (en) * | 1990-02-07 | 1996-01-10 | Ленинградский научно-исследовательский институт травматологии и ортопедии им.Р.Р.Вредена | Nutrient medium for clostridia isolation |
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