CN107365828A - Chromogenic culture medium for quick discriminating moraxelle catarrhalis - Google Patents
Chromogenic culture medium for quick discriminating moraxelle catarrhalis Download PDFInfo
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- CN107365828A CN107365828A CN201710872979.5A CN201710872979A CN107365828A CN 107365828 A CN107365828 A CN 107365828A CN 201710872979 A CN201710872979 A CN 201710872979A CN 107365828 A CN107365828 A CN 107365828A
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- moraxelle catarrhalis
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/045—Culture media therefor
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/21—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Pseudomonadaceae (F)
- G01N2333/212—Moraxellaceae, e.g. Acinetobacter, Moraxella, Oligella or Psychrobacter
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Abstract
The invention discloses a kind of chromogenic culture medium for quick discriminating moraxelle catarrhalis, including basal medium, 1 20 g/l of D glucose are also added with the basal medium, 5 50 mg/litre bromocresol purples, 50 500 mg/litre chromogenic substrates, 1 20 mg/litre X-factors and appropriate compound preservative, its pH value are 7.2 7.5.The advantage of the invention is that it accurately with the addition of a certain amount of chromogenic substrate and hemin in the basal medium that tradition uses, so that discrimination method is simple, without special installation during operation, also without extra bacteria distribution culture, separation differentiates to be carried out simultaneously, and the time is short and simple to operate, directly using sample, just energy quick separating identifies moraxelle catarrhalis, its sensitivity can reach 94.5%, and specificity reaches 95.1%, meets clinical detection and differentiates the demand of moraxelle catarrhalis.
Description
Technical field
The present invention relates to the culture medium of biological detection, more particularly, to a kind of for the aobvious of quick discriminating moraxelle catarrhalis
Color culture medium.
Background technology
Moraxelle catarrhalis（Moraxella catarrhalis）, once it is referred to as catarrh micrococcus luteus in the past（Micrococuus
catarrhalis）, mlicrococcus catarrhalis（Neisseria catarrhalis）, it is a kind of gram-negative diplococci,
Tympanitis, conjunctivitis, nasosinusitis, pneumonia, bronchitis, bacteremia and urinary system infection contamination etc. can be caused, the elderly and immune
The low crowd of power is susceptible.Foreign data shows that moraxelle catarrhalis is in Adult Lower Respiratory Tract Infections and children Streptococcus, tympanitis etc.
Separation rate in sample, is only second to haemophilus influenzae and streptococcus pneumonia, arranges the 3rd, moraxelle catarrhalis produces beta-lactam
The bacterial strain of enzyme is gradually more, and certain difficulty is brought for clinical treatment.Therefore rapidly and accurately detect and differentiate moraxelle catarrhalis, have than
Larger clinical practice meaning.
The existing detection and identification technology of moraxelle catarrhalis has following three kinds substantially：
1st, manual identification technology first uses broth medium flat board or blood agar culture-medium flat board or chocolate agar medium
Flat board etc. is separately cultured.Blood agar culture-medium flat board is such as used, 35 DEG C of culture 18-24 hours, moraxelle catarrhalis may have grown into light
Sliding, opaque, raised, canescence or white colony, this kind of doubtful bacterium colony of picking carry out a series of experiments and carry out discriminating checking：Such as
Bacterium colony is pushed away with oese to see and can promote, whether sugar fermentation is negative, if production hydrogen sulfide, whether form indoles, if reduction nitre
Whether hydrochlorate and nitrite, oxidizing ferment and catalase are positive, and whether DNA enzymatic is positive, if three butyronitrile of hydrolysis.Due to manual identification
Method not only grow by the time（Need two days）, and operation is extremely complex, and the preparation of a small amount of reagent in laboratory can waste substantial amounts of examination
Agent, directly purchases that complete indentifying substance price is higher and mass discrepancy is bigger, is as a result difficult to ensure.
2、Automating biochemical Bacteria Identification technical appraisement needs 24 hours, equally can not directly differentiate catarrh not from sample
Bacterium is drawn, the pure bacterium isolated can only be differentiated, and needs first to be separately cultured with plating medium, therefore the technology is reflected
Other moraxelle catarrhalis is also required to two days.The automation bacteria assessing instrument of the other technology costly, and identifies required mirror
Fixed card is also somewhat expensive.
3、Maldi-Tof MS（Matrix-assisted laser desorption ionization）Technology the method, which differentiates, to be taken
Between it is short, a few minutes can go out result, but can not directly differentiate moraxelle catarrhalis from sample, and need with plating medium it is advanced
Row is separately cultured, and the pure bacterium that isolated can only be differentiated, and being separately cultured for plating medium at least needs 1 day.Cause
Although this differentiates that the time is short, required time is not short, and Maldi-Tof MS instrument and equipments are sufficiently expensive, clinical labororatory
Equipment is also in the culture introduction period.
Chromogenic culture medium is that a kind of bacterium gradually risen the end of last century examines technology soon, is the spy having using bacterium
The opposite sex, by the method binding specificity zymolyte selectively cultivated, purpose bacterial growth and present is allowed to be grown with other
The visibly different color of bacterial clump or other colony characteristicses, so as to differentiate purpose bacterium.Colour developing culture biggest advantage is direct
Using sample, being separately cultured for bacterium is combined with discriminating, reaches the effect directly differentiated while separation.To mesh
Before untill, colour developing culture has had a variety of, such as Candida chromogenic medium, B group streptococcus chromogenic culture medium, golden yellow grape
Coccus chromogenic culture medium etc., but it is not yet found that moraxelle catarrhalis chromogenic culture medium comes out.
The content of the invention
It is an object of the invention to provide a kind of colour developing culture directly using sample with regard to energy quick discriminating moraxelle catarrhalis
Base.
To achieve the above object, the present invention can take following technical proposals：
Chromogenic culture medium of the present invention for quick discriminating moraxelle catarrhalis, including basal medium, on the basis
Also it is added with 1-20 g/l of D-Glucose in culture medium, 5-50 mg/litre bromocresol purples, 50-500 mg/litre chromogenic substrates,
1-20 mg/litres X-factor and appropriate compound preservative, its pH value are 7.2-7.5.
The compound preservative that the present invention uses includes 3 mg/litre vancomycins, 10 mg/litre nisins, 40,000
Units per liter polymyxin and 10 mg/litre nystatin.
Chromogenic substrate used in the present invention can use the chloro- 3- indoles-butyrates of 6-, can also use the chloro- 3- of the bromo- 6- of 5-
Indoles-butyrate, its content preferably 250 mg/litres.
In chromogenic culture medium of the present invention, preferably 5 g/l of the content of D-Glucose, preferably 10 milligrams of the content of bromocresol purple/
Rise, content preferably 5 mg/litres of X-factor.
The basal medium of the present invention includes yeast extract, peptone, beef liver leaching powder, sodium chloride, agar powder etc..
When prepared by chromogenic culture medium of the present invention, the composition of basal medium is claimed respectively by the formula of culture medium first
Amount, then absolute ethyl alcohol dissolving is added to prepare bromocresol purple solution, the chloro- 3- indoles-butyric acid salting liquids of 6- successively in proportion；It is hydrogenated with oxygen
Change sodium solution and prepare X-factor solution；After above-mentioned each raw material is mixed plus water carries out constant volume, and adjusts pH value and be
Sterilized after 7.2-7.5, the compound preservative added at 50 DEG C or so after being sterile filtered is cooled to, with sterile behaviour after being well mixed
The mode of work is filled into disposable plastic plate, and moraxelle catarrhalis chromogenic culture medium flat board is made.
In use, pressing Clinical microorganism laboratory procedures, directly sample streak inoculation to moraxelle catarrhalis is developed the color
On culture medium flat plate, 35 DEG C of culture 18-24 hours, you can according to the bacterial clump color grown on flat board（Aubergine bacterium colony）
Directly judge to differentiate in sample whether there is moraxelle catarrhalis.
The advantage of the invention is that accurately with the addition of in the basal medium that uses of tradition a certain amount of chromogenic substrate and
Hemin, so that discrimination method is simple, without special installation during operation, it is not required that extra bacteria distribution training
It is foster, separate discriminating and carries out simultaneously, the time is short and simple to operate, and directly identifying catarrh with regard to energy quick separating using sample does not draw
Bacterium, its sensitivity can reach 94.5%, and specificity reaches 95.1%, meet clinical detection and differentiate the demand of moraxelle catarrhalis.
Embodiment
Embodiment 1：Prepare 1 liter of culture medium
The formula of chromogenic culture medium of the present invention is：5 g/l of D-Glucoses, 10 mg/litre bromocresol purples, 250 mg/litre 6- are chloro-
3- indoles-butyrate, 5 mg/litre X-factors, 3 mg/litre vancomycins, 10 mg/litre nisins, 4
Ten thousand units per liter polymyxins, 10 mg/litre nystatin, 10 g/l of peptones, 3 g/l of yeast extracts, 4 g/l of beef liver leachings
Powder, 5 g/l of sodium chloride, 15 g/l of agar powders；
Load weighted 5 grams of D-Glucoses, 10 grams of peptones, 3 grams of yeast extracts, 4 grams of beef liver leaching powder and 5 grams of sodium chloride are added first
Suitable quantity of water dissolves together, the 10 mg/ml bromocresol purple solution and 2.5 then prepared by 1 milliliter of absolute ethyl alcohol dissolving is added
The chloro- 3- indoles-butyric acid salting liquids of 100 mg/ml 6- that milliliter absolute ethyl alcohol dissolving is prepared, add 1 milliliter of 0.1 equivalent hydroxide
The 5 mg/ml X-factor solution that sodium solution is prepared, add water to be settled to 1 liter, then add 15 grams of agar powders, and
Regulation pH value is sterilized after being 7.2-7.5, and it is stand-by that semi-finished product culture medium is obtained when being cooled to 50 DEG C or so；It is mould through the ages by 3 milligrams
Element, 9 milli liter of water of 10 milligrams of nisins and 40,000 unit polymyxins, then dissolve 10 with 1 milliliter of absolute ethyl alcohol
Milligram nystatin, is sterile filtered after the two is mixed, be then added to sterile manner cooled to the half of 50 DEG C or so into
In product culture medium, it is filled into after well mixed in a manner of sterile working in disposable plastic plate, moraxelle catarrhalis is made and shows
Color culture medium flat plate.
In use, pressing Clinical microorganism laboratory procedures, directly sample streak inoculation catarrh made of is not drawn
On bacterium chromogenic culture medium flat board, 35 DEG C of culture 18-24 hours, you can mirror is judged according to the bacterial clump color grown on flat board
Whether there is moraxelle catarrhalis in other sample：When the bacterial clump color grown on flat board is aubergine bacterium colony, it was demonstrated that sample
In contain moraxelle catarrhalis, when grown on flat board without bacterial clump or the bacterium that grows get blamed aubergine when, it was demonstrated that in sample
Without moraxelle catarrhalis.
Take 430 parts of clinical samples, including 285 parts of sputum specimen, bronchoalveolar lavage fluid 23, Urine specimens 79, purulent secretion
50 parts, 100 parts of leukorrhea sample, by more than on the 430 direct streak inoculation of sample oese to culture medium flat plates prepared above,
This 430 parts of samples are also seeded on blood agar plate simultaneously（Bacterium colony is separated on blood plate, is reflected for Brooker mass spectrograph
It is fixed）.Moraxelle catarrhalis chromogenic culture medium flat board after inoculation and blood agar plate are cultivated into 18-24 hours, picking for 35 DEG C simultaneously
Grown on the aubergine bacterium colony and blood agar plate grown on moraxelle catarrhalis chromogenic culture medium flat board smooth, impermeable
Bright, raised, canescence or white colony are identified simultaneously with Brooker Maldi-Tof MS, are identified and tied with Maldi-Tof MS
Fruit is defined.
It is positive 59 using Brooker Maldi-Tof MS qualification result display, it is negative 371；And use the present invention aobvious
Color culture medium flat plate identifies the moraxelle catarrhalis positive 56, negative 367.Statistical result showed：The inventive method it is sensitive
Up to 94.92%, specificity is fully able to meet clinical identification demand degree up to 98.92%.
Embodiment 2：Prepare 1 liter of culture medium
The formula of chromogenic culture medium of the present invention is：20 g/l of D-Glucoses, 30 mg/litre bromocresol purples, 400 mg/litre 6-
Chloro- 3- indoles-butyrate, 8 mg/litre X-factors, 3 mg/litre vancomycins, 10 mg/litre streptococcus lactis
Element, 40,000 units per liter polymyxins, 10 mg/litre nystatin, 10 g/l of peptones, 3 g/l of yeast extracts, 4 g/l of oxen
Liver soaks powder, 5 g/l of sodium chloride, 15 g/l of agar powders.
First by load weighted 20 grams of D-Glucoses, 10 grams of peptones, 3 grams of yeast extracts, 4 grams of beef liver leaching powder and 5 grams of chlorine
Changing sodium adds suitable quantity of water to dissolve together, the 10 mg/ml bromocresol purple solution then prepared by 1 milliliter of absolute ethyl alcohol dissolving is added
The chloro- 3- indoles-butyric acid salting liquids of 100 mg/ml 6- prepared with 4 milliliters of absolute ethyl alcohol dissolvings, add 1.6 milliliter of 0.1 equivalent
The 5 mg/ml X-factor solution that sodium hydroxide solution is prepared, add water to be settled to 1 liter, then add 15 grams of agar
Powder, and pH value is adjusted to be sterilized after 7.2-7.5, it is stand-by that semi-finished product culture medium is obtained when being cooled to 50 DEG C or so；3 milligram ten thousand
9 milli liter of water of ancient mycin, 10 milligrams of nisins and 40,000 unit polymyxins, then it is molten with 1 milliliter of absolute ethyl alcohol
10 milligrams of nystatin are solved, after the two is mixed, is sterile filtered, is then added to sterile manner and has cooled to 50 DEG C or so
In semi-finished product culture medium；It is filled into after well mixed in a manner of sterile working in disposable plastic plate, catarrh is made and does not draw
Bacterium chromogenic culture medium flat board.
Application method is the same as embodiment 1.
Embodiment 3：Prepare 1 liter of culture medium
The formula of chromogenic culture medium of the present invention is：10 g/l of D-Glucoses, 50 mg/litre bromocresol purples, 100 mg/litre 6-
Chloro- 3- indoles-butyrate, 18 mg/litre X-factors, 3 mg/litre vancomycins, 10 mg/litre streptococcus lactis
Element, 40,000 units per liter polymyxins, 10 mg/litre nystatin, 10 g/l of peptones, 3 g/l of yeast extracts, 4 g/l of oxen
Liver soaks powder, 5 g/l of sodium chloride, 15 g/l of agar powders.
First by load weighted 10 grams of D-Glucoses, 10 grams of peptones, 3 grams of yeast extracts, 4 grams of beef liver leaching powder and 5 grams of chlorine
Changing sodium adds suitable quantity of water to dissolve together, the 3 mg/ml bromocresol purple solution then prepared by 1 milliliter of absolute ethyl alcohol dissolving is added
The chloro- 3- indoles-butyric acid salting liquids of 100 mg/ml 6- prepared with 1 milliliter of absolute ethyl alcohol dissolving, add 1.4 milliliter of 0.1 equivalent
The 5 mg/ml X-factor solution that sodium hydroxide solution is prepared, add water to be settled to 1 liter, then add 15 grams of agar
Powder, and pH value is adjusted to be sterilized after 7.2-7.5, it is stand-by that semi-finished product culture medium is obtained when being cooled to 50 DEG C or so；3 milligram ten thousand
9 milli liter of water of ancient mycin, 10 milligrams of nisins and 40,000 unit polymyxins, then it is molten with 1 milliliter of absolute ethyl alcohol
10 milligrams of nystatin are solved, after the two is mixed, is sterile filtered, is then added to sterile manner and has cooled to 50 DEG C or so
In semi-finished product culture medium；It is filled into after well mixed in a manner of sterile working in disposable plastic plate, catarrh is made and does not draw
Bacterium chromogenic culture medium flat board.
Application method is the same as embodiment 1.
During actual preparation, the chloro- 3- indoles-butyrates of 6- can also replace with the chloro- 3- indoles-butyrates of the bromo- 6- of 5-, effect
Equivalent, equally, the raw material used in the present invention can carry out Reasonable adjustment in the value range of announcement, can meet to detect and differentiate
The clinical requirement of moraxelle catarrhalis.
Claims (5)
1. a kind of chromogenic culture medium for quick discriminating moraxelle catarrhalis, including basal medium, it is characterised in that：The base
It is added with 1-20 g/l of D-Glucose in basal culture medium, 5-50 mg/litre bromocresol purples, 50-500 mg/litre chromogenic substrates,
1-20 mg/litres X-factor and appropriate compound preservative, its pH value are 7.2-7.5.
2. the chromogenic culture medium according to claim 1 for quick discriminating moraxelle catarrhalis, it is characterised in that：It is described multiple
Closing bacteriostatic agent includes 3 mg/litre vancomycins, 10 mg/litre nisins, 40,000 units per liter polymyxins and 10 millis
G/l nystatin.
3. the chromogenic culture medium according to claim 1 for quick discriminating moraxelle catarrhalis, it is characterised in that：It is described aobvious
Color substrate is the chloro- 3- indoles-butyrates of 6- or the chloro- 3- indoles-butyrates of the bromo- 6- of 5-, and its content is 250 mg/litres.
4. the chromogenic culture medium according to claim 1 for quick discriminating moraxelle catarrhalis, it is characterised in that：The D-
The content of glucose is 5 g/l, and the content of the bromocresol purple is 10 mg/litres, and the content of the X-factor is
5 mg/litres.
5. the chromogenic culture medium according to claim 1 for quick discriminating moraxelle catarrhalis, it is characterised in that：The base
Basal culture medium includes yeast extract, peptone, beef liver leaching powder, sodium chloride, agar powder.
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| CN201710872979.5A CN107365828B (en) | 2017-09-25 | 2017-09-25 | Chromogenic medium for rapidly identifying moraxella catarrhalis |
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| CN201710872979.5A CN107365828B (en) | 2017-09-25 | 2017-09-25 | Chromogenic medium for rapidly identifying moraxella catarrhalis |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6130057A (en) * | 1999-09-28 | 2000-10-10 | Becton, Dickinson And Company | Method for differentiating microorganisms in a sample |
| EP1088896A2 (en) * | 1999-09-28 | 2001-04-04 | Becton Dickinson and Company | Chromogenic media containing blood or hemin |
| CN1298950A (en) * | 2000-07-14 | 2001-06-13 | 湖南省邵阳市中心医院 | Selective culture medium for cattamolar's bacteria |
| WO2005071096A2 (en) * | 2004-01-21 | 2005-08-04 | Molecular Probes, Inc. | Derivatives of cephalosporin and clavulanic acid for detecting beta-lacamase in a sample |
| CN105802891A (en) * | 2016-04-28 | 2016-07-27 | 山东鑫科生物科技股份有限公司 | Blood agar culture medium, disposable fastidious bacteria culture plate and preparation method thereof |
-
2017
- 2017-09-25 CN CN201710872979.5A patent/CN107365828B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6130057A (en) * | 1999-09-28 | 2000-10-10 | Becton, Dickinson And Company | Method for differentiating microorganisms in a sample |
| EP1088896A2 (en) * | 1999-09-28 | 2001-04-04 | Becton Dickinson and Company | Chromogenic media containing blood or hemin |
| CN1298950A (en) * | 2000-07-14 | 2001-06-13 | 湖南省邵阳市中心医院 | Selective culture medium for cattamolar's bacteria |
| WO2005071096A2 (en) * | 2004-01-21 | 2005-08-04 | Molecular Probes, Inc. | Derivatives of cephalosporin and clavulanic acid for detecting beta-lacamase in a sample |
| CN105802891A (en) * | 2016-04-28 | 2016-07-27 | 山东鑫科生物科技股份有限公司 | Blood agar culture medium, disposable fastidious bacteria culture plate and preparation method thereof |
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