CN201648404U - Novel two-phase culture medium of bacillus tuberculosis typus bovinus - Google Patents

Novel two-phase culture medium of bacillus tuberculosis typus bovinus Download PDF

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Publication number
CN201648404U
CN201648404U CN2010201749645U CN201020174964U CN201648404U CN 201648404 U CN201648404 U CN 201648404U CN 2010201749645 U CN2010201749645 U CN 2010201749645U CN 201020174964 U CN201020174964 U CN 201020174964U CN 201648404 U CN201648404 U CN 201648404U
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culture medium
bottle
sodium
ketopropionate
alpha
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Expired - Fee Related
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CN2010201749645U
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Chinese (zh)
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刘亚芹
吴长刚
杨振斌
冯冬霞
张法国
隋振亚
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Abstract

The utility model belongs to a medical supply, and provides a culture medium applicable to separation of bacillus tuberculosis typus bovinus. An outer bottle is a plastic bottle, and contains a sodium pyruvate solid culture medium inclined plane, a Sabouraud solid culture medium inclined plane and 10ml sodium pyruvate liquid culture medium; the height of the sodium pyruvate solid culture medium inclined plane is 45mm, the width of the part of the sodium pyruvate solid culture medium inclined plane at the bottom of the bottle is 15mm, and the opposite surface thereof is Sabouraud solid culture medium; the height of the Sabouraud solid culture medium is 45mm, and the width of the part of the Sabouraud solid culture medium at the bottom of the bottle is 15mm; a bottle cover is internally provided with a leak-proof rubber pad; and since the sodium pyruvate liquid culture medium can accelerate the growth of the bacillus tuberculosis typus bovinus and the Sabouraud culture medium can discover contaminants quickly, the culture period in check of the culture of the bacillus tuberculosis typus bovinus is shortened, and the working efficiency is improved.

Description

A kind of novel mycobacterium tuberculosis var bovis biphasic culture
Technical field
This novel mycobacterium tuberculosis var bovis biphasic culture belongs to a kind of medical treatment product.Having solved original substratum is the Sodium.alpha.-ketopropionate substratum for single solid medium only, and culture cycle is long, can not find the defective of contaminated bacteria fast.
Background technology
Clinically, the phlegm bacterium checks it is the most direct, the topmost foundation of diagnosis pulmonary tuberculosis and judgement pulmonary tuberculosis infectivity power, only turns out mycobacterium, could further do drug sensitive test and find the effective medicine of mycobacterium.Mycobacterium comprises mycobacterium tuberculosis complex and non-tuberculous mycobacteria.Comprise mycobacterium tuberculosis var bovis in the mycobacterium tuberculosis complex.The mycobacterium tuberculosis var bovis separation and Culture adopts the Sodium.alpha.-ketopropionate solid medium.
The Sodium.alpha.-ketopropionate solid medium:
Basal liquid Sodium Glutamate (purity is more than 99%) 7.2 grams
Potassium primary phosphate 2.4 grams
Sal epsom (MgSO 47H 2O) 0.24 gram
Magnesium citrate 0.6 gram
Sodium.alpha.-ketopropionate 1.6 grams
Glucose 4.0 grams
600 milliliters of distilled water
Inorganic salt and organism reagent all should adopt chemical pure (CP) reagent
1000 milliliters of fresh ovum gallinaceum liquid
20 milliliters of 2% malachite green solutions
The preparation method:
Measure 600 milliliters of distilled water.Add each inorganic salt composition, Sodium Glutamate and glycerol, 100 ℃ were heated 30 minutes, and the cooling back adds glucose.Clean fresh ovum gallinaceum surface, be immersed in 70% ethanol or other dilution thimerosal 20~30 minutes.Take out, dry, opening is collected ovum gallinaceum liquid, stirs evenly.Become ovum gallinaceum liquid by the disinfectant filtered through gauze.With 600 milliliters of basal liquids and 1000 milliliters of fresh ovum gallinaceum liquid mixings.Add 20 milliliters of 2% malachite green solutions, mixing, leave standstill 1 hour after, 7 milliliters of packing to master screw lid culture tube solidifies Sterilizers through substratum steam, 85 ℃ of sterilizations 50 minutes.
The preparation of Sodium.alpha.-ketopropionate liquid nutrient medium:
Aspartic acid 0.2g, MgSO 40.05g, Sodium.alpha.-ketopropionate 0.27g, KH 2PO 40.5g,, glucose 0.67g Sodium Citrate 0.5g, water 100ml, PH 7.2, add 2% malachite green solution 4ml, 121 ℃, add the aseptic blood plasma of 25ml after the high pressure cooling in 20 minutes.
The sabouraud culture medium preparation:
Peptone 10g, glucose 40g, agar powder 13g, water 1000ml, 121 ℃, sterile filling behind 20 minutes high pressure.
When doing the cultivation of mycobacterium tuberculosis var bovis of samples such as phlegm, body fluid, need carry out pre-treatment to it, promptly in centrifuge tube, add sample after, add sample volume 1-2 4% NaOH liquid vortex oscillation doubly again after 10 seconds, left standstill 20 minutes.
Require to improve the efficient that mycobacterium tuberculosis var bovis is cultivated clinically, what suitable mycobacterium tuberculosis var bovis was grown is the Sodium.alpha.-ketopropionate solid medium, in solid medium, mycobacterium tuberculosis var bovis grows up in three weeks of the bacterium colony needs above time, and gets final product in two weeks of fast growth in the Sodium.alpha.-ketopropionate liquid nutrient medium.Though pre-treatment can remove the impurity elimination bacterium as far as possible, also should reduce infringement simultaneously as far as possible to mycobacterium, therefore, pre-treatment can not remove the impurity elimination bacterium fully.In culturing process, also just inevitably there is contaminated bacteria, the contaminated bacteria great majority are fungies, in order to differentiate contaminated bacteria fast, add husky Bao Luoshi substratum again in the bottle of Sodium.alpha.-ketopropionate substratum is housed, sabouraud culture medium is to be exclusively used in the isolating substratum of fungus culture, and the fungi speed of growth thereon is very fast, so can find and differentiate contaminated bacteria fast, thereby improve culture efficiency.
Summary of the invention
The purpose of basal culture medium is to overcome above-mentioned problem, a kind of mycobacterium tuberculosis var bovis substratum that is applicable to is provided, on the basis of original Sodium.alpha.-ketopropionate solid medium, dispose Sodium.alpha.-ketopropionate liquid nutrient medium and husky Bao Luoshi substratum in the pipe simultaneously, the Sodium.alpha.-ketopropionate liquid nutrient medium is accelerated the growth of mycobacterium tuberculosis var bovis, sabouraud culture medium can be found contaminated bacteria fast, the shortening time in the inspection that mycobacterium tuberculosis var bovis is cultivated, raises the efficiency.The basal culture medium preparation process is the aseptic adding 10ml Sodium.alpha.-ketopropionate solid medium of elder generation, puts to add the 10ml sabouraud culture medium again after the inclined-plane cools off, and puts the inclined-plane cooling, and notice that two kinds of substratum do not mix, and add 10ml Sodium.alpha.-ketopropionate liquid nutrient medium at last this moment.
The technical solution of the utility model: a kind of novel mycobacterium tuberculosis var bovis biphasic culture is characterized in that: outer blank is a Plastic Bottle, the high 7cm of bottle, and the thick 1mm of bottle, bottle external diameter 4cm, the high 12mm of bottle cap covers thick 1mm, and leakproof sol pad is arranged in the bottle cap.In the bottle is Sodium.alpha.-ketopropionate culture medium slant, husky Bao Luoshi culture medium slant, 10ml Sodium.alpha.-ketopropionate liquid nutrient medium.The high 45mm of Sodium.alpha.-ketopropionate culture medium slant, the wide 15mm of the part at the bottom of bottle, the right opposite of Sodium.alpha.-ketopropionate solid medium are husky Bao Luoshi solid medium, the high 45mm of husky Bao Luoshi culture medium slant, the wide 15mm of the part at the bottom of bottle.Further technical scheme:
The sabouraud culture medium inclined-plane is arranged in the described device.
Dress Sodium.alpha.-ketopropionate liquid nutrient medium and Sodium.alpha.-ketopropionate solid medium in the described device.
The beneficial effects of the utility model are: this substratum can shorten the mycobacterium tuberculosis var bovis incubation time, and can find fast that contaminated bacteria has improved the quality inspection of mycobacterium tuberculosis var bovis.Show by large sample (520 example) experiment, use this substratum under identical condition, than simple Sodium.alpha.-ketopropionate solid medium cultivate speed fast a week, and finding and differentiating that the time of contaminated bacteria shortened to 2 days by original 5 to 6 days.After our unit uses this substratum, not only shorten the incubation time of mycobacterium tuberculosis var bovis, also reduced pollution rate, improved the culture efficiency of mycobacterium tuberculosis var bovis, be subjected to clinical doctor and the consistent of patient welcome.Having proved absolutely practicality, science, the characteristic of reasonable structure of this specimen bottle, is a kind of substratum of novel practical, is worth applying in mycobacterium tuberculosis var bovis cultivation check.
Description of drawings
The utility model is described in further detail below in conjunction with accompanying drawing:
Fig. 1 is the front view of the utility model substratum.
Fig. 2 is the left view of the utility model substratum.
Among the figure: 1 bottle cap, 2 bottles, 3 Sodium.alpha.-ketopropionate solid mediums, 4 sabouraud culture mediums, 5 Sodium.alpha.-ketopropionate liquid nutrient mediums
Embodiment:
As shown in Figure 1, 2, this mycobacterium tuberculosis var bovis culture apparatus is made up of culturing bottle and substratum, and culturing bottle is made for the PVC material, has the outer silk of spiral internal thread and bottleneck to cooperate, and bottle cap is made by silica gel, and bottle cap can closely be connected with bottle.In the bottle Sodium.alpha.-ketopropionate solid medium, Sodium.alpha.-ketopropionate liquid nutrient medium and sabouraud culture medium are housed.
The process of using this device to cultivate is:
(1) in refrigerator, takes out this culture apparatus.
Twist-off closure when (2) using, the low dip medium bottle, suction pipe after the use sterilization is drawn the liquid samples such as phlegm after the pre-treatment, add 2 earlier and cover the Sodium.alpha.-ketopropionate medium slant, after adding 2 covering sabouraud culture medium inclined-planes again, add 1 again and splash into the Sodium.alpha.-ketopropionate liquid nutrient medium, tighten bottle cap at last, vertically put 37 ℃ of cultivations.

Claims (2)

1. novel mycobacterium tuberculosis var bovis biphasic culture, be characterised in that: outer blank is a Plastic Bottle, the high 7cm of bottle, the thick 1mm of bottle, bottle external diameter 4cm, the high 12mm of bottle cap, cover thick 1mm, in the bottle is Sodium.alpha.-ketopropionate culture medium slant and husky Bao Luoshi culture medium slant and 10ml Sodium.alpha.-ketopropionate liquid nutrient medium, the high 45mm of Sodium.alpha.-ketopropionate culture medium slant, the wide 15mm of the part at the bottom of bottle, the right opposite of Sodium.alpha.-ketopropionate solid medium are husky Bao Luoshi solid medium, the high 45mm of husky Bao Luoshi culture medium slant, the wide 15mm of the part at the bottom of bottle.
2. according to right 1 described a kind of novel mycobacterium tuberculosis var bovis biphasic culture, leakproof sol pad is arranged in the bottle cap.
CN2010201749645U 2010-04-04 2010-04-04 Novel two-phase culture medium of bacillus tuberculosis typus bovinus Expired - Fee Related CN201648404U (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102191196A (en) * 2011-03-24 2011-09-21 湖南省天骑医学新技术有限公司 Mycobacterium tuberculosis solid/liquid culture medium and use method thereof
CN105145350A (en) * 2015-07-22 2015-12-16 青岛农业大学 Preparation method and application of PEG culture medium with solid-liquid two phases

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102191196A (en) * 2011-03-24 2011-09-21 湖南省天骑医学新技术有限公司 Mycobacterium tuberculosis solid/liquid culture medium and use method thereof
CN105145350A (en) * 2015-07-22 2015-12-16 青岛农业大学 Preparation method and application of PEG culture medium with solid-liquid two phases

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Granted publication date: 20101124

Termination date: 20110404