CN103397072B - Candida selective culture medium for feces specimen and preparation method thereof - Google Patents
Candida selective culture medium for feces specimen and preparation method thereof Download PDFInfo
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- CN103397072B CN103397072B CN201310289470.XA CN201310289470A CN103397072B CN 103397072 B CN103397072 B CN 103397072B CN 201310289470 A CN201310289470 A CN 201310289470A CN 103397072 B CN103397072 B CN 103397072B
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Abstract
The invention discloses a candida selective culture medium for a feces specimen and a preparation method thereof. The invention is characterized in that every 1000ml of culture medium contains malt extract powder, beef heart powder, magnesium sulfate, agar, gentamicin sulphate, aniline blue, calcium citrate tetrahydrate and the balance of distilled water. The preparation method comprises the following steps: dissolving the malt extract powder, beef heart powder, magnesium sulfate, agar, aniline blue, calcium citrate tetrahydrate, gentamicin sulphate, allithiamine and glycocholic acid in an orchis saina turcz decoction, evenly mixing, diluting to the specified volume, sterilizing and subpackaging. Compared with the prior art, the invention has the characteristics of high detection reliability and high candida separation rate.
Description
Technical field
Female case of the present invention is denomination of invention: a kind of candidiasis Selective agar medium for stool sample and preparation method thereof, application number 2012100564838, the applying date: the division on March 6th, 2012, relate to inspection microorganism culturing field, be specifically related to a kind of candidiasis Selective agar medium for stool sample.
Background technology
Microorganism culturing is a kind of technology making microbial growth by artificial means, and the substratum that Selective agar medium is used to promote or suppress the organism (as cell or bacterium etc.) of certain type and designs, utilize this substratum required microorganism can be separated from the microorganism mixed.Candidiasis is modal conditioned pathogen in fungi, and also known as candiyeast, it often parasitizes the places such as the skin of people, oral cavity, vagina and intestinal mucosa, when human body immune function lowly or is normally lived away from home the micro-ecological environment imbalance at position, easily causes moniliosis.The Clinical Laboratory means mainly Gram staining of current candidiasis, but have the candidiasis Gram staining of 20% positive to detect.And the kind of candidiasis is differentiated as determined, just need to cultivate, and differentiate with regard to its colony morphology characteristic.But the candidiasis that current clinical Mycophyta Selective agar medium used is mainly used in the clinical samples such as urine, phlegm, blood, bile, ascites is separated, as potato dextrose agar (PDA) substratum, low for the candidiasis separation rate in stool sample, its reason is that the intestinal bacteria in ight soil etc. are faster than candidiasis growth on current fungi culture medium more, cover substratum very soon, candidiasis cannot grow.
Summary of the invention
Technical assignment of the present invention is for above the deficiencies in the prior art, provides a kind of stool sample that can be used in, the Selective agar medium high to candidiasis separation rate.
The technical scheme that the present invention solves its technical problem is: a kind of candidiasis Selective agar medium for stool sample, and its feature exists, and contains in the formula of preparation 1000ml substratum:
Fructus Hordei Germinatus leaching powder 3 ~ 10g;
OX-heart powder 10 ~ 20g;
Magnesium sulfate 1.2 ~ 2g;
Agar 15 ~ 20g;
Gentamicin sulphate 0.005 ~ 0.02g;
Aniline orchid 0.1 ~ 0.4g;
Four water citric acid calcium 0.01 ~ 0.2g;
Distilled water adds to 1000ml.
In prioritization scheme, every 1000ml substratum is also containing allithiamine 0.05 ~ 0.5g.
In prioritization scheme, every 1000ml substratum is also containing glycocholic acid 0.01 ~ 0.1g.
In prioritization scheme, the Herba Orchidis Latifoliae Aqueous extracts 200 ~ 300ml of every 1000ml substratum also containing concentration 200g/L.
For a preparation method for the candidiasis Selective agar medium of stool sample, it is characterized in that comprising the following steps:
(1) get Herba Orchidis Latifoliae and add water boil 0.5 ~ 1 hour, filter, get filtrate, adjustment volume is 200g/L to concentration, obtains Herba Orchidis Latifoliae Aqueous extracts;
(2) get Fructus Hordei Germinatus leaching powder, OX-heart powder, magnesium sulfate, agar, aniline orchid, four water citric acid calcium, gentamicin sulphate, allithiamine, glycocholic acid be dissolved in step 1 gained Herba Orchidis Latifoliae Aqueous extracts, mixing, distilled water constant volume, 121 DEG C, packing after 15min sterilizing.
Wherein said Fructus Hordei Germinatus leaching powder, OX-heart powder provide carbon nitrogen source and energy; Magnesium sulfate is buffer reagent, and can destroy antimycotic rhzomorph residual in sample; Agar is the peptizer of substratum; Gentamicin sulphate suppresses varied bacteria growing, improves the separation rate of candidiasis; Aniline orchid is indicator; Four water citric acid calcium are buffer reagent and activator; Allithiamine is nutrition-fortifying agent, can promote glucose uptake; Glycocholic acid suppresses Gram-positive bacteria growing; Herba Orchidis Latifoliae is the herb of orchid Mongolia Herba Orchidis Latifoliae, and herb contains carbohydrate 90%, protein 2.5%, fat 0.4%, and 80% in carbohydrate is phlegmatic temperament and starch, the free sugar of 20% and robust fibre; Semi-lactosi is got, fructose, seminose, wood sugar and galacturonic acid in phlegmatic temperament; Raffinose is had, maltose, sucrose, glucose and wood sugar in free sugar; Be rich in Methionin, α-amino-isovaleric acid and palmitinic acid in protein at interior 15 kinds of lipid acid, its Aqueous extracts is good nutrition-fortifying agent, and has the effect suppressing intestinal bacilli growth.
The present invention compared with prior art, has and detects the feature reliable, candidiasis separation rate is high.
Embodiment
Below in conjunction with practical situation, the specific embodiment of the present invention is elaborated.
Embodiment 1, contains in the formula of preparation 1000ml substratum: Fructus Hordei Germinatus leaching powder 3g; OX-heart powder 20g; Magnesium sulfate 1.2g; Agar 15g; Gentamicin sulphate 0.02g; The blue 0.1g of aniline; Four water citric acid calcium 0.2g; Distilled water adds to 1000ml.Preparation method: get Fructus Hordei Germinatus leaching powder, OX-heart powder, magnesium sulfate, agar, aniline orchid, four water citric acid calcium, gentamicin sulphate be dissolved in distilled water, mix, distilled water is settled to 1000ml, 121 DEG C, packing after 15min sterilizing.
Embodiment 2, contains in the formula of preparation 1000ml substratum: Fructus Hordei Germinatus leaching powder 10g; OX-heart powder 10g; Magnesium sulfate 2g; Agar 20g; Gentamicin sulphate 0.005g; The blue 0.4g of aniline; Four water citric acid calcium 0.01g; Distilled water adds to 1000ml.Preparation method: get Fructus Hordei Germinatus leaching powder, OX-heart powder, magnesium sulfate, agar, aniline orchid, four water citric acid calcium, gentamicin sulphate be dissolved in distilled water, mix, distilled water is settled to 1000ml, 121 DEG C, packing after 15min sterilizing.
Embodiment 3, contains in the formula of preparation 1000ml substratum: Fructus Hordei Germinatus leaching powder 3g; OX-heart powder 20g; Magnesium sulfate 1.2g; Agar 15g; Gentamicin sulphate 0.02g; The blue 0.1g of aniline; Four water citric acid calcium 0.2g, allithiamine 0.05g, glycocholic acid 0.1g; Distilled water adds to 1000ml.Preparation method: get Fructus Hordei Germinatus leaching powder, OX-heart powder, magnesium sulfate, agar, aniline orchid, four water citric acid calcium, gentamicin sulphate, allithiamine, glycocholic acid be dissolved in distilled water, mix, distilled water is settled to 1000ml, 121 DEG C, packing after 15min sterilizing.
Embodiment 4, contains in the formula of preparation 1000ml substratum: Fructus Hordei Germinatus leaching powder 10g; OX-heart powder 10g; Magnesium sulfate 2g; Agar 20g; Gentamicin sulphate 0.005g; The blue 0.4g of aniline; Four water citric acid calcium 0.01g, allithiamine 0.5g, glycocholic acid 0.01g; Distilled water adds to 1000ml.Preparation method: get Fructus Hordei Germinatus leaching powder, OX-heart powder, magnesium sulfate, agar, aniline orchid, four water citric acid calcium, gentamicin sulphate, allithiamine, glycocholic acid be dissolved in distilled water, mix, distilled water is settled to 1000ml, 121 DEG C, packing after 15min sterilizing.
Embodiment 5, contains in the formula of preparation 1000ml substratum: Fructus Hordei Germinatus leaching powder 6g; OX-heart powder 15g; Magnesium sulfate 1.5g; Agar 18g; Gentamicin sulphate 0.01g; The blue 0.3g of aniline; Four water citric acid calcium 0.1g, allithiamine 0.3g, glycocholic acid 0.05g; Distilled water adds to 1000ml.Preparation method: get Fructus Hordei Germinatus leaching powder, OX-heart powder, magnesium sulfate, agar, aniline orchid, four water citric acid calcium, gentamicin sulphate, allithiamine, glycocholic acid be dissolved in distilled water, mix, distilled water is settled to 1000ml, 121 DEG C, packing after 15min sterilizing.
Embodiment 6, contains in the formula of preparation 1000ml substratum: Fructus Hordei Germinatus leaching powder 3g; OX-heart powder 20g; Magnesium sulfate 1.2g; Agar 15g; Gentamicin sulphate 0.02g; The blue 0.1g of aniline; Four water citric acid calcium 0.2g, allithiamine 0.05g, the Herba Orchidis Latifoliae Aqueous extracts 200ml of glycocholic acid 0.1g, concentration 200g/L; Distilled water adds to 1000ml.Preparation method: (1) is got 40g Herba Orchidis Latifoliae and added water boil 0.5 hour, filters, gets filtrate, according to filtrate volume, taking the concentrated or adjustment volume that adds water to be 200g/L to concentration, obtaining Herba Orchidis Latifoliae Aqueous extracts 200ml; (2) get Fructus Hordei Germinatus leaching powder, OX-heart powder, magnesium sulfate, agar, aniline orchid, four water citric acid calcium, gentamicin sulphate, allithiamine, glycocholic acid be dissolved in step 1 gained Herba Orchidis Latifoliae Aqueous extracts, mixing, distilled water is settled to 1000ml, 121 DEG C, packing after 15min sterilizing.
Embodiment 7, contains in the formula of preparation 1000ml substratum: Fructus Hordei Germinatus leaching powder 10g; OX-heart powder 10g; Magnesium sulfate 2g; Agar 20g; Gentamicin sulphate 0.005g; The blue 0.4g of aniline; Four water citric acid calcium 0.01g, allithiamine 0.5g, the Herba Orchidis Latifoliae Aqueous extracts 300ml of glycocholic acid 0.01g, concentration 200g/L; Distilled water adds to 1000ml.Preparation method: (1) is got 60g Herba Orchidis Latifoliae and added water boil 1 hour, filters, gets filtrate, and adjustment volume is 200g/L to concentration, obtains Herba Orchidis Latifoliae Aqueous extracts 300ml; (2) get Fructus Hordei Germinatus leaching powder, OX-heart powder, magnesium sulfate, agar, aniline orchid, four water citric acid calcium, gentamicin sulphate, allithiamine, glycocholic acid be dissolved in step 1 gained Herba Orchidis Latifoliae Aqueous extracts, mixing, distilled water is settled to 1000ml, 121 DEG C, packing after 15min sterilizing.
Embodiment 8, contains in the formula of preparation 1000ml substratum: Fructus Hordei Germinatus leaching powder 6g; OX-heart powder 15g; Magnesium sulfate 1.5g; Agar 18g; Gentamicin sulphate 0.01g; The blue 0.3g of aniline; Four water citric acid calcium 0.1g, allithiamine 0.3g, the Herba Orchidis Latifoliae Aqueous extracts 250ml of glycocholic acid 0.05g, concentration 200g/L; Distilled water adds to 1000ml.Preparation method: (1) is got 50g Herba Orchidis Latifoliae and added water boil 0.8 hour, filters, gets filtrate, and adjustment volume is 200g/L to concentration, obtains Herba Orchidis Latifoliae Aqueous extracts 300ml; (2) get Fructus Hordei Germinatus leaching powder, OX-heart powder, magnesium sulfate, agar, aniline orchid, four water citric acid calcium, gentamicin sulphate, allithiamine, glycocholic acid be dissolved in step 1 gained Herba Orchidis Latifoliae Aqueous extracts, mixing, distilled water is settled to 1000ml, 121 DEG C, packing after 15min sterilizing.
Gained candidiasis Selective agar medium of the present invention is used for stool sample to be had and detects the feature reliable, candidiasis separation rate is high, and be clinical data sufficient proof, pertinent data is as follows.
1 object and method.
1.1 medium preparing: establish 4 groups altogether, be respectively embodiment 8 scheme group, embodiment 5 scheme group, embodiment 2 scheme group and control group, control group is potato dextrose agar (PDA) substratum.
1.2 cultural methods: gather outpatient service or Hospitalized Diarrhea patient fresh excreta, be suspended in 5ml0.9% physiological saline, with transfering loop sectional streak in separating plate, cultivate 24h for 37 DEG C, the tiny bacterium colony of picking carries out identification of bacteria.
1.3 statistical analysis SPSS13.0 carry out statistical study, and P<0.05 indicates significant.
2 results: at 969 parts of stool samples, be separated and be accredited as candidiasis, embodiment 8 scheme group is 167 strains, embodiment 5 scheme group is 158 strains, embodiment 2 scheme group is 155 strains, control group PDA substratum is 124 strains, and through statistical procedures, various embodiments of the present invention group compares with control group and has notable difference (embodiment 8 scheme group P=0.006; Embodiment 5 scheme group P=0.029; Embodiment 2 scheme group P=0.045).Result shows, embodiment of the present invention candidiasis separation rate is the high existing PDA substratum being usually used in fungus culture obviously.
Claims (1)
1. for a preparation method for the candidiasis Selective agar medium of stool sample, it is characterized in that: Fructus Hordei Germinatus leaching powder 6g; OX-heart powder 15g; Magnesium sulfate 1.5g; Agar 18g; Gentamicin sulphate 0.01g; The blue 0.3g of aniline; Four water citric acid calcium 0.1g, allithiamine 0.3g, the Herba Orchidis Latifoliae Aqueous extracts 250ml of glycocholic acid 0.05g, concentration 200g/L; Distilled water adds to 1000ml; Preparation method comprises the following steps:
(1) get Herba Orchidis Latifoliae and add water boil 0.5 ~ 1 hour, filter, get filtrate, adjustment volume is 200g/L to concentration, obtains Herba Orchidis Latifoliae Aqueous extracts;
(2) get Fructus Hordei Germinatus leaching powder, OX-heart powder, magnesium sulfate, agar, aniline orchid, four water citric acid calcium, gentamicin sulphate, allithiamine, glycocholic acid be dissolved in step 1 gained Herba Orchidis Latifoliae Aqueous extracts, mixing, distilled water constant volume, 121 DEG C, packing after 15min sterilizing.
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| CN101948902A (en) * | 2010-09-10 | 2011-01-19 | 中国检验检疫科学研究院 | Candida chromogenic medium, detection kit and detection method |
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| 李斯特菌的选择培养基和分离方法;杨其智等;《中国人兽共患病杂志》;19921231;第8卷(第1期);第52、53页 * |
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