CN100345977C - Preparing method for cow mammitis staphylococcus culture fluid and its use - Google Patents
Preparing method for cow mammitis staphylococcus culture fluid and its use Download PDFInfo
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Abstract
The present invention relates to a preparation method and a purpose for milk cow mammitis staphylococcus culture fluid, which belongs to the field of an animal epidemic disease diagnose and control technology. 10g of peptone, 5g of yeast powder, 10g of sodium pyruvate, 12g of glycine and 5g of lithium chloride are fully dissolved in 980 ml of water, sodium hydroxide solution is used to regulate a pH value to 7.3, then sterilization is carried out under the condition of 121 DEG C, cooling is carried out until a temperature is 50 DEG C (+/-)10 DEG C, 10 ml of sterile potassium tellurite with a concentration of 1 %, and 7 ml of tween 80 are added, and preservation is carried out as a temperature of 4 DEG C. Prepared culture fluid can be used for rapid milk cow mammitis staphylococcus identification and a mammitis staphylococcus drug sensitive test.
Description
Technical field
The invention belongs to animal epidemic diagnosis and Prevention Technique field.
Technical background
Mammitis of cow sickness rate height, hazardness is big, is one of main disease that influences world's dairy development.According to the data record of international milk federation, nearly 50% cow suffers from mastitis, and wherein nearly 2% suffers from clinic mastitis, and 48% suffers from latent mammitis.China's clinic mastitis sickness rate 33.41% (9.7%～55.6%), the recall rate of latent mammitis are 50-80%.Clinic mastitis has lactation amount decline, mammary swelling, milk significantly clinical symptom such as rotten, though and the breast of latent mammitis and milk outward appearance do not have the visible naked eyes and change, usually cause the quality of raw material milk to descend, threaten that people's is healthy.According to estimates, China every year because of financial loss that mastitis caused above 3.16 hundred million yuan.The pathogenic micro-organism that causes mastitis mainly comprises gold-coloured staphylococci, streptococcus agalactiae, streptococcus dysgalactiae, cow's milk room suis, intestinal bacteria, klebsiella, Pseudomonas aeruginosa and some other fungi, virus etc.But in these pathogenic agent,, account for whole mammitis of cow case more than 90% based on streptococcus aureus, suis and intestinal bacteria.
The treatment of mammitis of cow is based on microbiotic, but because antibiotic use lack of standardization has caused serious problems such as antibiotic remains in the milk, Resistant strain is general, the mastitis that pasts medical help case increases.The major cause that causes this phenomenon is because the restriction of diary farm economy, technology and experiment condition, seldom carrying out necessary bacterium before with antibiotic control mammitis of cow separates and drug sensitivity test, only depend on animal doctor's hobby and experience, long-term, blindly and excess use antibiotic.
Domestic and international normally used drug sensitive test method mainly contains disk diffusion method, broth dilution method, agar dilution, automatization susceptibility instrument method and Etest method at present.Wherein, disk diffusion method is the responsive or tolerance of judging bacterial antibiotic by the diameter of measuring inhibition zone, great advantage is that expense is low and medicament selection is flexible, inoculated multiple factor affecting such as bacterium amount, incubation time, antibiotic content and diffusive force, plate thickness but exist, need strictness to carry out stdn, just can guarantee result's reliability, and have consuming time, effort and shortcomings such as inapplicable anerobe and severe oxygen bacterium; Broth dilution method and agar dilution are the methods of the microbiotic doubling dilution being come quantitative assay minimum inhibitory concentration (MIC) in meat soup or agar, its advantage is can accurately measure some anerobes and severe oxygen bacterium to antibiotic susceptibility, and one the cover test tube can survey many strains bacterium, but have shortcoming such as consuming time and effort; Automatization susceptibility instrument is to come sentence read result by detecting turbidity, the fluorescence intensity of fluorescent indicator or the hydrolysis reaction of fluorogenic substrate, have quick and advantage such as powerful function of statistic analysis, lack handiness, be difficult to detect shortcomings such as some special bacterium and instrument cost an arm and a leg but have medicament selection; The Etest method is in conjunction with the advantage of diffusion process and dilution method, with the susceptibility that contains continuous concentration gradient microbiotic plastic strip bacterial detection, has accurately, advantage such as reliable, good stability, can be used for the drug sensitive test of various bacteria, and unique shortcoming is to cost an arm and a leg.
Drug sensitive test for cow mammitis staphylococcus, method commonly used both at home and abroad at present is the milk sample inoculation sheep blood agar plate with aseptic collection, cultivate after 18-48 hour for 37 ℃ and make preliminary evaluation according to colonial morphology and haemolysis characteristics, the suspicious bacterium colony of picking carries out biochemical tests such as Thrombin coagulase and confirms again, use colonies typical on Mueller-Hinton agar plates such as (MH), to carry out the paper disk method drug sensitive test at last, or carry out the mensuration of MIC with broth dilution method.These methods not only needed just can obtain a result at least in 3-5 days, possessed equipment, reagent and technical qualification that necessary bacteriology is identified, and can not truly reflect sometimes that with the drug sensitive test result that single bacterium colony carries out bacterium " colony " is to antibiotic susceptibility.As for automatization susceptibility instrument method and Etest method, be difficult to by domestic veterinary clinic and diary farm acceptance because price is very expensive.
Summary of the invention
Purpose of the present invention provides a kind of compound method of the cow mammitis staphylococcus culture fluid that can be fast staphylococcus selectivity in the milk milk sample be spread cultivation.
The present invention is dissolved in 10g peptone, 5g yeast powder, 10g Sodium.alpha.-ketopropionate, 12g glycine, 5g lithium chloride in the 980ml water earlier fully, transfer pH to 7.3 with sodium hydroxide solution again, then, in 121 ℃ of sterilizations down, add aseptic 1% potassium tellurite solution 10ml and 7ml tween 80 (Tween-80) when being cooled to 50 ℃ ± 10 ℃, 4 ℃ of preservations.
Another purpose provides the purposes of above-mentioned cow mammitis staphylococcus culture fluid among the present invention:
One of purposes is: the method that is used to differentiate cow mammitis staphylococcus: the nutrient solution that aforesaid method is obtained is sub-packed in the cultivation plate hole, inoculation milk sample, cultivated 24 hours under 37 ℃ of conditions, black product and muddy appears producing in nutrient solution, and judging in the milk sample has staphylococcus.
Two of purposes is: be used for the drug sensitive test of cow mammitis staphylococcus: be coated with the nutrient solution that the antibiotic cultivation plate hole adding of international standard MIC aforesaid method obtains, inoculate the milk sample, under 35 ℃ of conditions, cultivated 24 hours, measure staphylococcus antibiotic susceptibility.
The present invention selects to cultivate on the Baird-Parker medium base in staphylococcus, replace extractum carnis and yeast extract paste with yeast powder, replace yolk liquid with tween 80 (Tween-80), be and be the transparence staphylococcus culture fluid, called after mBP nutrient solution, be the mBP agar plate as adding 1.5% agar powder again, can be used for cow mammitis staphylococcus and select fast/differentiate.Can not only guarantee the inhibition of the nutritional need and the non-aureus growth of aureus growth, and make nutrient solution transparent, so that whether muddy and produce the black product and judge and have or not aureus growth according to nutrient solution after the microbial culture.
When having or not staphylococcus in judging the milk sample, after cultivating 24 hours, black product and muddiness appear in nutrient solution, then, in the sample of can clearly suckling staphylococcus are arranged.As nutrient solution still is transparent liquid, then, does not have staphylococcus in the milk sample.
When measuring staphylococcus, after cultivating 24 hours, whether produce the black product according to nutrient solution and have or not aureus growth to reach corresponding antibiotic susceptibility or resistance with muddy judgement to antibiotic susceptibility.
The present invention selects on the basis of substratum in existing staphylococcus, by improvement to culture medium prescription and cultural method, develop the method that directly to carry out quick discriminating of staphylococcus and drug sensitive test with clinical milk sample, its remarkable advantage is quick, accurate, inexpensive and does not need complicated plant and instrument, is particularly suitable for domestic middle-size and small-size diary farm and the animal doctor of basic unit and uses.
Description of drawings
Fig. 1 is the growth curve chart of streptococcus aureus in mBP nutrient solution and ordinary broth.
Specific embodiment
One, preparation mBP nutrient solution:
Take by weighing peptone 10g, yeast powder 5g, Sodium.alpha.-ketopropionate 10g, glycine 12g, lithium chloride 5g respectively in Glass Containers with balance, add water 980ml, stir and fully the dissolving after, transfer pH to 7.3 with sodium hydroxide solution, sterilization is 15 minutes under 121 ℃ of high temperature, add the 1% potassium tellurite solution 10ml and the 7ml tween 80 (Tween-80) of filtration sterilization when being cooled to 50 ℃ ± 10 ℃, 4 ℃ of preservations are standby then.
Two, staphylococcic quick discriminating:
Judge tentatively according to clinical examination or California mastitis test (CMT) detected result and to suffer from clinical type or latent mammitis milk cow, squeeze with conventional method of milking and remove first three milk that usefulness routine disinfection liquid disinfectant nipple and near zone thereof are with the cotton ball soaked in alcohol nipple of sterilizing.The sterilised test tube lid is opened, on the other hand test tube is kept horizontality, another hand milking, and milk " is injected " in the test tube, and build the test tube lid immediately, place ice bath to take back real (inspection) immediately and test the chamber.In aseptic worktable, open 24 or 96 aseptic well culture plates, the mBP nutrient solution is divided in each hole of culture plate, every hole 200 μ l, in every hole, add undiluted or with the suitable milk sample 10 μ l of dilution of physiological saline, build back 37 ℃ and leave standstill and cultivated 24 hours, whether muddy and produce the black product and judge and have or not aureus growth according to nutrient solution.
Three, staphylococcic quick medicine-sensitive test:
1, determining and culture plate bag quilt of antibiotic concentration: with reference to the disk diffusion method bacteriostatic test method and the judging criterion of the stdn council of international standard U.S. clinical labororatory (NCCLS20012) recommendation, filter out the some strains of streptococcus aureus to examination antibiotic sensitivity, it is carried out pure culture on the plain agar flat board, the single colony inoculation 2ml of picking nutrient broth is cultivated after 12 hours for 37 ℃ and is measured usefulness for MIC respectively.0.5,1,2 or 3 times of microbiotic to standard MIC, 200 μ l mBP nutrient solutions and 10 μ l staphylococcus cultures are added in each hole of 96 well culture plates, cultivate after 24 hours for 35 ℃, determine the microbiotic MIC that drug sensitive test is required according to no responsive aureus growth (the not muddy and no black product of nutrient solution).Microbiotic is made into 1000 times of MIC after proofreading and correct, is sub-packed in aseptic each hole of 96 well culture plates, in aseptic worktable, dry up or lyophilize after, 4 ℃ of preservations are standby.
2, staphylococcic quick medicine-sensitive test: in aseptic worktable, the mBP nutrient solution is sub-packed in is coated with antibiotic 96 well culture plates, every hole 200 μ l, in each hole, add 10 μ l staphylococcuses respectively and select culture or mastitis milk sample for the first time, cultivated 24 hours for 35 ℃, whether muddy and produce the black product and judge and have or not aureus growth and whether responsive according to nutrient solution to corresponding microbiotic.
Three, simultaneous test:
1, the growing state of streptococcus aureus in the mBP substratum: 25 strains are suffered from the isolating streptococcus aureus streak inoculation of ox mBP agar plate from mastitis, cultivate after 24 hours for 37 ℃, bacterial growth becomes typical staphylococcus bacterium colony, diameter is 0.5-1.5mm, the edge is translucent, central black.The picking colonies typical is made nacterial smear, and microscopy behind gramstaining is typical thyrsiform.The picking colonies typical carries out biochemical test, and 25 strain bacteriums all show as clotting of plasma enzyme positive, the catalase positive, and 7.5% high salt growth is positive.
Picking streptococcus aureus colonies typical inoculation nutrient broth was cultivated after 24 hours for 37 ℃, and the ratio inoculation mBP nutrient solution with 1% in 37 ℃ of culturing process, was got one time sample, with spectrophotometric instrumentation OD every 2 hours
600Value is with OD
600Value is ordinate zou, and incubation time is that X-coordinate is described growth curve.As can be seen, the growth of streptococcus aureus in the mBP nutrient solution enters logarithmic phase dynamically to similar substantially in nutrient broth after 4 hours, enters the growth platform phase in 14 hours from the growth curve of bacteria (Fig. 1), growth rate decline after 24 hours.
2, the selectivity characteristic of mBP substratum: the nutrient broth that respectively streptococcus aureus, staphylococcus epidermidis, Bacillus subtilus, bacillus rhusiopathiae suis, suis, intestinal bacteria, Salmonellas, Bacillus proteus, pasteurella multocida, Pseudomonas aeruginosa, shigella dysenteriae inoculation is contained 5% calf serum, cultivate after 24 hours for 37 ℃, get 100 μ l cultures respectively and coat the mBP agar plate, cultivated 24 hours for 37 ℃, except that streptococcus aureus, other bacterium there is no visible growth.Get the nutrient broth culture 20 μ l of above-mentioned bacterium respectively, inoculation is gone in the 2ml mBP nutrient solution, cultivate after 24 hours for 37 ℃, except that the streptococcus aureus inoculated tube, the color and the opacity of all the other microbionation pipe nutrient solutions all do not change, and show that only streptococcus aureus can grow in the mBP nutrient solution.
20 μ l streptococcus aureus broth cultures, 200 μ l intestinal bacteria broth cultures and suis broth culture are mixed, get 40 μ l and inoculate nutrient solution into 4ml mBP, cultivate after 24 hours for 37 ℃, get 100 μ l cultures coating suis and intestinal bacteria respectively and select substratum, cultivate after 48 hours for 37 ℃, do not see intestinal bacteria and streptococcus growth, prove that further the mBP nutrient solution has very strong restraining effect to intestinal bacteria and streptococcus growth.
The growth curve of streptococcus aureus in mBP nutrient solution and ordinary broth as shown in Figure 1, among the figure, ordinate zou is the absorbance value of bacterial cultures, X-coordinate be incubation time (hour); The growth curve of selective presentation streptococcus aureus in quick selection/discriminating mBP nutrient solution; The growth curve of common expression streptococcus aureus in nutrient broth.
3, separate staphylococcic sensitivity with the mBP nutrient solution:, with physiological saline streptococcus aureus nutrient broth culture is carried out continuous 10 times of dilutions, until 10 with 1: 9 ratio
-8, from every pipe, draw 50 μ l and inoculate nutrient solution into 1ml mBP, from last three pipes, draw 100 μ l coating nutrient agar medium, cultivate after 24 hours 1-7 pipe (10 for 37 ℃
-1-10
-7) in all have muddy and the black product, aureus growth has been described, enumeration result demonstration, it is 60 bacteriums that the mBP nutrient solution separates staphylococcic sensitivity.
4, the mBP nutrient solution is to staphylococcic separating effect in the clinical milk sample: detect (CMT method) result according to clinical diagnosis and milk sample somatocyte, 4 parts in clinical type of aseptic collection and recessive mammitis of cow milk sample, sheep blood agar plate partition method and biochemical test with routine are defined as infection of staphylococcus aureus milk sample, it is suitably inoculated mBP nutrient solution, mBP agar plate and nutrient agar plate respectively after the dilution, cultivate after 24 hours for 37 ℃, in the mBP nutrient solution staphylococcus aureus growth is arranged, culture is obviously muddy, and the black product is arranged; The enumeration result shows that the streptococcus aureus colony number on mBP agar plate and the nutrient agar plate is near (table 1).
5, mBP nutrient solution composition is to the influence of microbiotic bacteriostatic activity: use streptococcus aureus 7 strains that are separated to from clinical milk sample, with reference to NCCLS20012 Staphylococcus disk diffusion method drug sensitive test method and judging criterion, on the MH substratum, carry out drug sensitive test, determine that 7 strain streptococcus aureuses are to 78 kinds of antibiotic susceptibility of class (table 2).Above-mentioned streptococcus aureus is cultivated on the plain agar flat board, and the single colony inoculation 2ml of picking nutrient broth is cultivated after 12 hours for 37 ℃ and is carried out 96 well culture plate method drug sensitive tests respectively.With mBP nutrient solution packing 96 well culture plates, every hole 200 μ l, 0.5,1,2 or 3 times of antibiotic to standard MIC is added in the respective aperture, 10 μ l streptococcus aureus nutrient broth cultures are inoculated in every hole, cultivate after 24 hours for 35 ℃, whether muddy and produce the black product and judge and have or not aureus growth and according to nutrient solution to corresponding antibiotic susceptibility.As shown in table 3, with the mBP nutrient solution, the drug sensitive test result who carries out on 96 well culture plates with 8 kinds of microbiotic of 7 classes of 4 concentration is consistent with the disk diffusion method result of standard, shows that mBP nutrient solution composition does not have obvious influence to the antibiotic fungistatic effect of try.
6, carry out the quick medicine-sensitive test with the mBP nutrient solution: according to clinical examination and milk sample somatocyte CMT detected result, gather 7 parts in the positive milk of mastitis sample, every part of milk sample is divided into two, is respectively applied for the disk diffusion method drug sensitive test and is the quick medicine-sensitive test of nutrient solution with mBP.The mBP nutrient solution is sub-packed in is coated with antibiotic 96 orifice plates of standard MIC, every hole 200 μ l, in each hole, add the clinical milk sample of 10 μ l respectively, cultivate after 24 hours for 35 ℃, whether muddy and produce the black product and judge and have or not aureus growth and to corresponding antibiotic susceptibility, the result shows two kinds of drug sensitive test methods and results unanimities (table 4) according to nutrient solution.
Above test-results shows: mBP nutrient solution (base) has very strong selection specificity, sensitivity and distinguishing ability to the cow mammitis streptococcus aureus.With mBP nutrient solution and 24 or 96 hole microbial culture plate combinations, set up the method for quick identification of cow mammitis staphylococcus, whether as long as nutrient solution is sub-packed in cultivates plate hole, inoculation trace milk sample and cultivate through 24 hours, can clearly draw is the conclusion of staphylococcal infections.To be coated with the combination of antibiotic 96 well culture plates of international standard MIC and mBP nutrient solution, set up the quick medicine-sensitive test method of cow mammitis staphylococcus, as long as with nutrient solution packing, inoculation micro-example and cultivation in 24 hours, can know clearly that staphylococcus is to which kind of antibiotic sensitive or resistance, not only can be used for the drug sensitive test of staphylococcus pure culture, the more important thing is and directly to carry out drug sensitive test with clinical milk sample, its outstanding advantage is quick, easy, inexpensive and does not need specialized equipment equipment, is particularly suitable for diary farm and clinical animal doctor and uses.
Table 1: from clinical milk sample, separate staphylococcic effect relatively with different substratum
Annotate: +++expression milk sample CMT detected result is the strong positive latent mammitis; + expression aureus growth the positive.
Table 2 international standard NCCLS20012 is about the explanation of drug sensitive test of streptococcus aureus disk diffusion method and relevant MIC
S represents responsive, and I represents medium sensitivity, and R represents insensitive (resistance).
Table 3:7 strain streptococcus aureus is to 7 kinds of antibiotic susceptibility (standard disk diffusion method)
Annotate: with reference to NCCLS20012 Staphylococcus drug sensitive test method and judging criterion as a result, S represent sensitivity, and I represents medium sensitivity, and R represents insensitive (resistance).
Table 4: standard disk diffusion method and mBP liquid culture method streptococcus aureus drug sensitive test result's comparison
| Microbiotic | Bacterial strain | Paper disk method | The MQQ liquid culture method | |||
| 0.5MIC | 1.0MIC | 2.0MIC | 3.0MIC | |||
| Penicillin | 1184 | R(22) | + | + | + | + |
| 1416 | S(38) | - | - | - | - | |
| 1094 | R(21) | + | + | + | + | |
| 1084 | R(10) | + | + | + | + | |
| Clindamycin | 1416 | S(25) | - | - | - | - |
| 1184 | S(29) | - | - | - | - | |
| 1084 | S(24) | - | - | - | - | |
| 0002 | R(13) | + | + | + | + | |
| Ceftriaxone | 1184 | S(27) | - | - | - | - |
| 0002 | S(29) | - | - | - | - | |
| 1084 | S(33) | - | - | - | - | |
| 9938 | I(20) | + | + | + | + | |
| Compound sulfanilamide (SN) | 1084 | R(7) | + | + | + | + |
| 9730 | R(9) | + | + | + | + | |
| 1184 | R(7) | + | + | + | + | |
| 1094 | R(8) | + | + | + | + | |
| Ciprofloxacin | 1416 | S(24) | - | - | - | - |
| 1184 | I(19) | + | + | + | + | |
| 1084 | S(24) | - | - | - | - | |
| 9730 | I(18) | + | + | + | + | |
| Erythromycin | 1416 | S(24) | - | - | - | - |
| 1084 | S(25) | - | - | - | - | |
| 0002 | R(9) | + | + | + | + | |
| 1094 | R(8) | + | + | + | + | |
| Tsiklomitsin | 1094 | I(13) | + | + | + | + |
| 9938 | R(11) | + | + | + | + | |
| 1416 | S(17) | - | - | - | - | |
| 0002 | S(23) | - | - | - | - | |
Annotate: 0.5,1.0,2.0,3.0 multiples that are respectively standard MIC, S, I, R represent respectively bacterial antibiotic in the disk diffusion method drug sensitive test sensitivity, in quick and resistance; "+" and "-" represents respectively in containing the antibiotic mBP nutrient solution of different concns or does not have bacterial growth, judges bacterial antibiotic resistance or sensitivity thus.
MBP liquid culture method and standard disk diffusion method drug sensitive test result that table 5 carries out with clinical milk sample and streptococcus aureus pure culture
| Strain number | Method | Microbiotic | ||||||
| Penicillin | Clindamycin | Ceftriaxone | Compound sulfanilamide (SN) | Ciprofloxacin | Erythromycin | Tsiklomitsin | ||
| 9730 | Paper disk method | R | S | S | R | S | R | R |
| The mBP/ pure culture | + | - | - | + | - | + | + | |
| The mBP/ sample of suckling | + | - | - | + | - | + | + | |
| 9938 | Paper disk method | R | S | I | R | S | S | R |
| The mBP/ pure culture | + | - | - | + | - | - | + | |
| The mBP/ sample of suckling | + | - | - | + | - | - | + | |
| 1094 | Paper disk method | R | R | I | R | S | R | R |
| The mBP/ pure culture | + | + | + | + | - | + | + | |
| The mBP/ sample of suckling | + | + | + | + | - | + | + | |
| 1184 | Paper disk method | R | S | S | R | S | S | S |
| The mBP/ pure culture | + | - | - | + | - | - | - | |
| The mBP/ sample of suckling | + | - | - | + | - | - | - | |
| 1416 | Paper disk method | S | S | S | R | S | S | S |
| The mBP/ pure culture | + | - | - | + | - | - | - | |
| The mBP/ sample of suckling | + | - | - | + | - | - | - | |
| 0002 | Paper disk method | R | R | S | R | - | R | R |
| The mBP/ pure culture | + | + | - | + | - | + | + | |
| The mBP/ sample of suckling | + | + | - | + | - | + | + | |
| 1084 | Paper disk method | R | S | S | R | S | S | R |
| The mBP/ pure culture | + | - | - | + | - | - | + | |
| The mBP/ sample of suckling | + | - | - | + | - | - | + | |
S, R represent bacterial antibiotic sensitivity and resistance in the disk diffusion method drug sensitive test respectively; "+" and "-" represents respectively in containing the antibiotic mBP nutrient solution of standard MIC or does not have aureus growth, judges bacterial antibiotic resistance or sensitivity thus.
Claims (3)
1, a kind of compound method of cow mammitis staphylococcus culture fluid, it is characterized in that: earlier 10g peptone, 5g yeast powder, 10g Sodium.alpha.-ketopropionate, 12g glycine, 5g lithium chloride are dissolved in the 980ml water fully, transfer pH to 7.3 with sodium hydroxide solution again, then, in 121 ℃ of sterilizations down, add aseptic 1% potassium tellurite solution 10ml and 7ml tween 80 when being cooled to 50 ℃ ± 10 ℃, 4 ℃ of preservations.
2, the cow mammitis staphylococcus culture fluid according to the described method preparation of claim 1 is used to differentiate the staphylococcic method of mammitis of cow, it is characterized in that: nutrient solution is sub-packed in the cultivation plate hole, inoculation milk sample, under 37 ℃ of conditions, cultivated 24 hours, black product and muddiness appear in nutrient solution, and judging in the milk sample has staphylococcus.
3, the purposes that is used for the cow mammitis staphylococcus drug sensitive test according to the cow mammitis staphylococcus culture fluid of the described method preparation of claim 1, it is characterized in that: in being coated with the antibiotic cultivation plate hole of international standard MIC, add nutrient solution, inoculate the milk sample, under 35 ℃ of conditions, cultivated 24 hours, measure staphylococcus antibiotic susceptibility.
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| CN1236813A (en) * | 1998-04-27 | 1999-12-01 | 齐齐哈尔市卫生防疫站 | Culture medium for quick separation and identification of staphylococcus |
| CN1369550A (en) * | 2001-02-16 | 2002-09-18 | 沈阳协合集团有限公司 | Culture medium of staphylococcus aureus and its preparing process |
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| CN1236813A (en) * | 1998-04-27 | 1999-12-01 | 齐齐哈尔市卫生防疫站 | Culture medium for quick separation and identification of staphylococcus |
| CN1369550A (en) * | 2001-02-16 | 2002-09-18 | 沈阳协合集团有限公司 | Culture medium of staphylococcus aureus and its preparing process |
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