CN104845917A - Enrichment culture medium - Google Patents

Enrichment culture medium Download PDF

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Publication number
CN104845917A
CN104845917A CN201510298137.4A CN201510298137A CN104845917A CN 104845917 A CN104845917 A CN 104845917A CN 201510298137 A CN201510298137 A CN 201510298137A CN 104845917 A CN104845917 A CN 104845917A
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powder
culture medium
sterilizing
vitamin
bacterium
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钟学文
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    • 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)
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  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
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  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention discloses an enrichment culture medium and belongs to the field of detection. The enrichment culture medium is characterized in that a formula contains beef liver powder, malt extract powder, yeast extract powder, sodium chloride, agar, ferric ammonium citrate, calcium lactate, D-seromycin, 2-ethoxymethylene-3,5-dihydroxy-y-pyrone, vitamin K1, sterile defiberized sheep blood and distilled water. Compared with the prior art, the enrichment culture medium plays roles in transporting and enriching.

Description

A kind of enrichment medium
Technical field
The present invention relates to field of medical examination, be specifically related to a kind of enrichment medium.
Background technology
It is by the chronic disease of the bacterial a kind of Zoonosis of Nocardia that slave blocks bacterium disease (Norcardosis), is characterized as tissue suppuration, downright bad or formation abscess.This disease is distributed widely in all over the world, and using pulmonary nocardiosis as common, slave blocks bacterium and causes the primary of people, suppurative pulmonary infection by respiratory tract, can occur phthisical symptom.Pulmonary lesions can be transferred to subcutis, forms abscess, ulcer and multiple fistula, also can be diffused into other organs.Similar to pulmonary tuberculosis, pulmonary aspergillosis, lung Wegner granuloma in clinical symptom, be easy to mistaken diagnosis.
Clinical diagnosis many dependence microbial culture of this disease, concrete grammar is: have visible colonies after cultivating 3 days with common blood agar culture plate (BAP), and after 7 ~ 10 days, bacterium colony is protruding, and after aerial hyphae is formed, surface is in fine hair shape.Existing training method there is following problem: 1. because nocadia belongs to severe bacteria, namely harsher to growing environment, nutritional requirement bacterium, growth or can not be difficult in conventional environment, therefore reliable diagnosis needs just can complete for 7 days, often delay and clinically the Accurate Diagnosis of bacterium disease is blocked to slave and takes effective remedy measures in time, and the case fatality rate causing slave to block bacterium disease increases; 2., in collection of specimens and transport process, due to varied bacteria growing, it is suppressed that slave blocks bacterium, and cause slave block bacterium separation and Culture positive rate on the low side on whole slave block bacterium disease diagnosis impact huge.Therefore, guarantee fully increase bacterium in transport process and prevent living contaminants very important.
Summary of the invention
Technical assignment of the present invention is for above the deficiencies in the prior art, provides a kind of mycetoma diagnosis substratum that slave blocks bacterium positive rate of can guaranteeing.
The technical scheme that the present invention solves its technical problem is: a kind of enrichment medium, and its feature exists, and contains in often liter: beef liver powder 10.0g; Fructus Hordei Germinatus leaching powder 2.0g; Yeast extract powder 5.0g; Sodium-chlor 5.0g; Agar 15.0g; Ferric Ammonium Citrate 0.1g; Calcium lactate 0.05g; D-Cycloserine 0.05g; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone 0.001g; Vitamin K 10.1g; Aseptic defiber Sheep Blood 100ml; ; Distilled water adds to 1000ml.
The preparation method of above-mentioned substratum is: take beef liver powder; Fructus Hordei Germinatus leaching powder; Yeast extract powder; Sodium-chlor; Agar; Ferric Ammonium Citrate; Calcium lactate; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone mixes, and distilled water dissolves, and 210 DEG C, sterilizing 15min, is cooled to 45 DEG C, adds sterilizing D-Cycloserine; Sterilizing vitamin k 1; Sterilizing network protein phosphatase polypeptide; Aseptic defiber Sheep Blood mixing, packing after sterile purified water constant volume to 1000ml.
Wherein: described beef liver powder, Fructus Hordei Germinatus leaching powder provides carbon source, nitrogenous source, the nutritive ingredients such as yeast extract powder rich in proteins, amino acid, polypeptide, Nucleotide, VITAMIN, somatomedin, trace element, for microorganism provides balanced nutritious, these materials are to promoting that the Growth and Reproduction that slave blocks bacterium is all very important factor; Sodium-chlor maintains balanced osmotic pressure; Agar is the peptizer of substratum; Ferric Ammonium Citrate is nutrition agent, and for slave blocks the ferro element that bacterium flourish provides required, and under this concentration, Ferric Ammonium Citrate also has buffer reagent effect; Calcium lactate, except doing nutrient intensifier, can also suppress gram-positive cocci to grow; While D-Cycloserine provides nutrition, under this concentration, also suppress the growth of Gram-negative bacteria; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone (2-Ethoxymethylene-3,5-dihydroxy-y-pyrone), laboratory study shows that 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone can suppress the growth of the miscellaneous bacterias such as staphylococcus, suis, Bacillus proteus, bacillus enteritidis, multiple fungi, but under this concentration due to Nocardia without antibacterial and bacteriostasis, therefore improve the separation rate that slave blocks bacterium.By experiment same slave being blocked bacterium is inoculated on 2 BAP substratum, and one of them BAP substratum adds 0.1% sterilizing vitamin k 1cultivate at 37 DEG C, after 48 hours, observe bacterial strain, find to add vitamin k 1bAP substratum have little slave to block bacterium bacterium colony, do not add vitamin k 1bAP substratum do not find that slave blocks bacterium bacterium colony, vitamin k 1promote cell's growth, differentiation are the positive growth factors that slave blocks bacterium, contribute to shortening incubation time; Aseptic defiber Sheep Blood provides energy environment; Phosphopeptide caseinate is activator, can promote that slave blocks the picked-up of bacterium to ferro element, calcium constituent; L-asparagine is nutrition-fortifying agent, promotes that slave blocks bacterium to amino acid whose picked-up; Radix Rehmanniae is the fresh of goatweed Radix Rehmanniae or dried root, and modern medicine study shows, containing more than 20 kind of glycoside in Radix Rehmanniae rhizome; 8 kinds of carbohydrates; More than 20 seed amino acids and more than 20 plant inorganic elements, and Radix Rehmanniae Aqueous extracts is good nutrition-fortifying agent, and have the effect suppressing staphylococcus aureus growth.
The present invention compared with prior art, has and detects reliable, slave and block the high feature of bacterium separation rate.
Embodiment
Below in conjunction with practical situation, the specific embodiment of the present invention is elaborated.
Embodiment 1, filling a prescription is: beef liver powder 10.0g; Fructus Hordei Germinatus leaching powder 2.0g; Yeast extract powder 5.0g; Sodium-chlor 5.0g; Agar 15.0g; Ferric Ammonium Citrate 0.1g; Calcium lactate 0.05g; D-Cycloserine 0.05g; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone 0.001g; Vitamin K 10.1g; Aseptic defiber Sheep Blood 100ml; ; Distilled water adds to 1000ml.Preparation method takes beef liver powder; Fructus Hordei Germinatus leaching powder; Yeast extract powder; Sodium-chlor; Agar; Ferric Ammonium Citrate; Calcium lactate; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone mixing distilled water dissolves, and 210 DEG C, sterilizing 15min, is cooled to 45 DEG C, adds sterilizing sterilizing D-Cycloserine; Sterilizing vitamin K 1; Aseptic defiber Sheep Blood mixing, packing after sterile purified water constant volume to 1000ml.
Embodiment 2, filling a prescription is: beef liver powder 10.0g; Fructus Hordei Germinatus leaching powder 5.0g; Yeast extract powder 5.0g; Sodium-chlor 5.0g; Agar 15.0g; Ferric Ammonium Citrate 0.3g; Calcium lactate 0.03g; D-Cycloserine 0.03g; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone 0.003g; Vitamin K 10.01g; Network protein phosphatase polypeptide 0.1g; Aseptic defiber Sheep Blood 80ml; ; Distilled water adds to 1000ml.Preparation method takes beef liver powder; Fructus Hordei Germinatus leaching powder; Yeast extract powder; Sodium-chlor; Agar; Ferric Ammonium Citrate; Calcium lactate; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone mixes, and distilled water dissolves, and 210 DEG C, sterilizing 15min, is cooled to 45 DEG C, adds sterilizing sterilizing D-Cycloserine; Sterilizing vitamin k 1; Sterilizing network protein phosphatase polypeptide; Aseptic defiber Sheep Blood mixing, packing after sterile purified water constant volume to 1000ml.
Embodiment 3, filling a prescription is: beef liver powder 5.0g; Fructus Hordei Germinatus leaching powder 3.0g; Yeast extract powder 10.0g; Sodium-chlor 5.0g; Agar 20.0g; Ferric Ammonium Citrate 0.2g; Calcium lactate 0.03g; D-Cycloserine 0.01g; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone 0.001g; Vitamin K 10.03g; Network protein phosphatase polypeptide 0.3g; Aseptic defiber Sheep Blood 90ml; The Radix Rehmanniae Aqueous extracts 250ml of concentration 200g/L; ; Distilled water adds to 1000ml.Preparation method preparation method: (1) is got 50g Radix Rehmanniae and added water boil 45min, filters, gets filtrate, according to filtrate volume, taking the concentrated or adjustment volume that adds water to be 200g/L to concentration, obtaining Radix Rehmanniae Aqueous extracts 250ml; (2) beef liver powder is taken; Fructus Hordei Germinatus leaching powder; Yeast extract powder; Sodium-chlor; Agar; Ferric Ammonium Citrate; Calcium lactate; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone mixing is dissolved in the Radix Rehmanniae extracting solution of step (1) gained and mixes, and 210 DEG C of sterilizing 15min, are cooled to 45 DEG C, add sterilizing sterilizing D-Cycloserine; Sterilizing network protein phosphatase polypeptide; Sterilizing vitamin K 1; Aseptic defiber Sheep Blood mixing, packing after sterile purified water constant volume to 1000ml.
Embodiment 4, filling a prescription is: beef liver powder 8.0g; Fructus Hordei Germinatus leaching powder 3.0g; Yeast extract powder 8.0g; Sodium-chlor 5.0g; Agar 20.0g; Ferric Ammonium Citrate 0.1g; Calcium lactate 0.01g; D-Cycloserine 0.05g; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone 0.005g; Vitamin K 10.05g; Network protein phosphatase polypeptide 0.3g; L-asparagine 0.01g; Aseptic defiber Sheep Blood 100ml; The Radix Rehmanniae Aqueous extracts 300ml of concentration 200g/L; ; Distilled water adds to 1000ml.Preparation method: (1) is got 60g Radix Rehmanniae and added water boil 60min, filters, gets filtrate, according to filtrate volume, taking the concentrated or adjustment volume that adds water to be 200g/L to concentration, obtaining Radix Rehmanniae Aqueous extracts 300ml; (2) beef liver powder is taken; Fructus Hordei Germinatus leaching powder; Yeast extract powder; Sodium-chlor; Agar; Ferric Ammonium Citrate; Calcium lactate; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone; L-asparagine mixing is dissolved in the Radix Rehmanniae extracting solution of step (1) gained and mixes, and 210 DEG C of sterilizing 15min, are cooled to 45 DEG C, add sterilizing sterilizing D-Cycloserine; Sterilizing network protein phosphatase polypeptide; Sterilizing vitamin K 1; Aseptic defiber Sheep Blood mixing, packing after sterile purified water constant volume to 1000ml.Described Radix Rehmanniae Aqueous extracts concentration 200g/L refers to often liter containing crude drug 200g.
Clinical trial data is as follows.
Setting control group and experimental group, described control group uses common blood agar culture plate (BAP) substratum, and its formula is: beef powder 5g/L, peptone 12g/L, agar 15g/L, sodium-chlor 5g/L, aseptic defiber Sheep Blood 100ml/L.Described experimental group A, B, C, D corresponding embodiment 1 scheme group respectively; Embodiment 2 scheme group, embodiment 3 scheme group, embodiment 4 scheme group.
The highly doubtful slave of clinical acquisitions 29 example blocks bacterium patient sample (phlegm, segmental bronchus extractum, fester etc.), and each increment originally adds in five groupings respectively, and containing two identical substratum in each grouping, is labeled as respectively " transhipment " and " transhipment ".The processing scheme of " transhipment " is: leave simulation in clinical laboratory's censorship vehicle in room temperature (25 DEG C) situation and transport environment, get back to clinical laboratory and carry out microbial culture (35 DEG C) after 24h; The processing scheme of " not transporting " is carry out microbial culture (35 DEG C) after sample collecting immediately.The microbial culture environmental parameter of all samples is consistent, and the tiny bacterium colony of each picking of 4d, 7d carries out identification of bacteria all after incubation.Positive bacterial culture detects standard and complys with " clinical microbiology diagnosis and diagram ".
After five groups of microbial culture, after 4d, 7d, the positive detects and the results are shown in following table,
Result shows: compare between group during the microbial culture 4d 1. not transporting and transport, and embodiment 1 ~ 4 and control group positive rate all exist huge difference (embodiment 1 group of P<0.05, embodiment 2 ~ 4 groups of P<0.01); And during 7d, do not transport group group difference not obvious (P>0.05), transhipment group embodiment 1 ~ 4 positive rate is apparently higher than control group (P<0.05); 2., in embodiment 1 ~ 4 group, transport and do not transport positive rate no significant difference (P>0.05), and control group two time points compare, obvious difference (P<0.05).
Can draw, the severe jamming using BAP cannot resist the temperature of transhipment and condition target Pseudomonas grow, and substratum of the present invention can keep object bacteria suitably to grow, thus the separation of raising object bacteria.

Claims (1)

1. an enrichment medium, is characterized in that, contains in often liter: beef liver powder 10.0g; Fructus Hordei Germinatus leaching powder 2.0g; Yeast extract powder 5.0g; Sodium-chlor 5.0g; Agar 15.0g; Ferric Ammonium Citrate 0.1g; Calcium lactate 0.05g; D-Cycloserine 0.05g; 2-Ethoxymethylene-3,5-dihydroxy-y-pyrone 0.001g; Vitamin K 10.1g; Aseptic defiber Sheep Blood 100ml; ; Distilled water adds to 1000ml.
CN201510298137.4A 2015-06-04 2015-06-04 Enrichment culture medium Pending CN104845917A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107400642A (en) * 2017-07-24 2017-11-28 内蒙古东达獭兔循环产业研究院 A kind of bacteriotrophy culture medium and preparation method thereof
CN111248026A (en) * 2020-04-07 2020-06-09 中国科学院沈阳应用生态研究所 Quercus matsutake culture medium and application thereof
CN111607535A (en) * 2020-05-13 2020-09-01 南京农业大学 Application of cycloserine and beneficial bacteria in cooperation for preventing and controlling soil-borne bacterial wilt of tomatoes

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107400642A (en) * 2017-07-24 2017-11-28 内蒙古东达獭兔循环产业研究院 A kind of bacteriotrophy culture medium and preparation method thereof
CN111248026A (en) * 2020-04-07 2020-06-09 中国科学院沈阳应用生态研究所 Quercus matsutake culture medium and application thereof
CN111248026B (en) * 2020-04-07 2022-04-08 中国科学院沈阳应用生态研究所 Quercus matsutake culture medium and application thereof
CN111607535A (en) * 2020-05-13 2020-09-01 南京农业大学 Application of cycloserine and beneficial bacteria in cooperation for preventing and controlling soil-borne bacterial wilt of tomatoes
CN111607535B (en) * 2020-05-13 2021-02-09 南京农业大学 Application of cycloserine and beneficial bacteria in cooperation for preventing and controlling soil-borne bacterial wilt of tomatoes

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Application publication date: 20150819