CN101633899B - Fast-growing solid culture medium of tubercle bacillus and standardized production method thereof - Google Patents
Fast-growing solid culture medium of tubercle bacillus and standardized production method thereof Download PDFInfo
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Abstract
The invention relates to a fast-growing solid culture medium of tubercle bacillus, comprising solution A and solution B, wherein the A solution includes: 1.0-2.0g of monopotassium phosphate, 0.3-0.5g of magnesium sulfate, 0.5-0.8g of magnesium citrate, 2.0-3.0g of asparagine, 15-20g of agar powder and 800ml of deionized water; the solution B comprises: 3.0-4.0g of bovine serum albumin, 4-5g of glucose, 1.2-1.8g of catalase, 5-10g of oleic acid, 0.01-0.10g of triphosadenine, 0.3-0.8g of coenzyme A, 0.03-0.09g of cytochrome C, 0.06-0.10g of Alpha-naphthylacetic acid, 600Mu of polymyxin B, 60Mug of amphotercin B, 240Mug of nalidixic acid, 60Mug of trimethoprim, 60Mug of azlocillin, and 200ml of deionized water; more preferably, the A solution also comprises 0.02-0.03g of ammonium ferric citrate, edible gourmet powder for replacing asparagine and human or animal strum for replacing bovine serum albumin; the invention also provides a standardized production method of the fast-growing solid culture medium. The fast-growing solid culture medium of the tubercle bacillus has smart design, simple and convenient preparation, low cost, stable and reliable stable, easy standardized large-scale production and fast-growing tubercle bacillus cultured on the medium, thereby ensuring a drug sensitive test to be fast and accurate and being good for early diagnosis and treatment of the tubercle bacillus.
Description
Technical field
The present invention relates to the biology techniques field, particularly tubercule bacillus culture technique field specifically is meant a kind of fast-growing solid culture medium of tubercle bacillus and standardized production method thereof.
Background technology
For a long time, laboratory diagnosis lungy mainly relies on the bacteriology smear and cultivates inspection.Because method is limit, and has had a strong impact on timely diagnosis lungy.Therefore, for many years Chinese scholars seeking fast always, responsive, special, easy laboratory diagnostic method.
One, smear and cultivation
Smear is the most basic bacteriology checking method of white plaque with cultivating.The sensitivity of cultivating is a little more than smear method, and is the reliable method of identifying viable bacteria, also can further carry out strain identification and drug sensitive test, is described as " golden standard " of diagnosis of tuberculosis.But conventional Russell medium incubation time is long, need 4-8 week just can go out the result, and positive rate also has only 30-40%; Various mycobacteriums all can grow simultaneously, need to combine strain identification just can determine whether to be Mycobacterium tuberculosis.Because the positive rate of smear and cultivation is on the low side, causes a large amount of tuberculosis patients to be failed to pinpoint a disease in diagnosis or mistaken diagnosis, has had a strong impact on the needs of clinical diagnosis and treatment.In addition, solid culture Quito commonly used now is basic nutrition source with egg, because egg source difference causes composition uncertain, unsuitable industrialization is difficult to stdn.
Two, fast culture, susceptibility detection technique
Mycobacterium fast culture, the susceptibility detection technique that be born the seventies in 20th century are a kind of new white plaque bacteriology checking methods.With Bactec TB 460 is first-generation mycobacterium fast culture, the susceptibility detection system of representative, and quick diagnosis lungy has been played vital role.Numerous application results shows that Bactec TB 460 can significantly shorten incubation time (being coated with yang disease example average positive fate 9-14 days); Drug sensitive test only needs 5-7 days; Simultaneously positive rate improves 10% approximately, and easy and simple to handle, robotization is strong, also can carry out quick bacterial type and identify.But the subject matter that exists is can't observe colonial morphology in the liquid nutrient medium, and pollution rate is higher than Luo Shi and cultivates, and instrument more expensive with the reagent price, radiocontamination is arranged, so be difficult to the all-round popularization application.
With Bactec MGIT 960 is s-generation mycobacterium fast culture, the susceptibility detection system of representative; Solved the radioactive pollution that Bactec TB460 exists; And kept its advantage fast, white plaque bacteriology checking method has been had again further develop.This cultivation appearance can be used for phlegm, segmental bronchus washing liquid, hydrothorax, ascites, cerebrospinal fluid, urine, fester, synovial fluid, lesion tissue or the mycobacterium cultivation of the various clinical samples of cheesy masses, preliminary strain identification and drug sensitivity test.External application result shows, the cultured positive rate of Bactec MGIT 960, positive time, pollution rate are all close with Bactec TB 460.But the level of automation of Bactec MGIT 960 is higher, the operation is more easy, and shortcoming is that price is higher.
The required condition of the growth and breeding of tubercule bacillus is higher, in its substratum the multiple nutrients material need be arranged, and the kind of nutritive substance is few in the existing solid tubercule bacillus substratum; Make tubercule bacillus in existing solid tubercule bacillus substratum, can not carry out growth and breeding well like this, production rate is slow, and mitotic cycle reaches 12-16 hour; Want 4-6 week can see bacterium colony; Therefore, use existing solid tubercule bacillus substratum to come tubercule bacillus is carried out external artificial separation when cultivating, its detection speed is quite slow; Positive rate is low, is unfavorable for the early diagnosis and therapy of tubercule bacillus.In addition; Traditional tuberculosis drug sensitive test is that antitubercular agent is joined in the Luo Shi solid medium by certain extent of dilution; The tubercule bacillus that then separation and Culture is gone out is inoculated in the substratum that contains medicine, observes the growing state of tubercule bacillus in substratum and judges.Because growth of bacillus tubercle is slow, this traditional method needs the time in 4-6 week could report the drug sensitive test result usually, so can't in time for clinical rational drug use guidance be provided.Therefore the time that shortens the tubercule bacillus drug sensitive test has become a problem that presses for solution, and quick, sensitive tuberculosis drug sensitive test then is the most important condition of controlling tuberculosis.
Owing to there is above-mentioned defective, therefore, need a kind of new method cultivation tubercule bacillus of adopting, make the tubercule bacillus growth fast,, thereby make drug sensitive test fast, accurately, help the early diagnosis and therapy of tubercule bacillus.
Summary of the invention
The objective of the invention is to have overcome above-mentioned shortcoming of the prior art; A kind of fast-growing solid culture medium of tubercle bacillus and standardized production method thereof are provided; The design of this fast-growing solid culture medium of tubercle bacillus is ingenious, simple and convenient preparation, cost is low, steady quality is reliable, be easy to stdn scale operation; The tubercule bacillus growth of cultivating above that is quick, thereby makes drug sensitive test fast, accurately, helps the early diagnosis and therapy of tubercule bacillus.
To achieve these goals,, fast-growing solid culture medium of tubercle bacillus is provided, has been characterized in, having comprised in first aspect of the present invention:
A liquid: 1.0-2.0g potassium primary phosphate (KH
2PO
4), 0.3-0.5g sal epsom (MgSO
4), 0.5-0.8g magnesium citrate, 2.0-3.0g l-asparagine and 15-20g agar powder, the 800ml deionized water;
B liquid: 3.0-4.0g bovine serum albumin (bovine albumin), 4-5g glucose (dextrose), 1.2-1.8g katalase (catalase), 5-10g oleic acid (oleic acid), 0.01-0.10g Triphosaden (ATP), 0.3-0.8g coenzyme A, 0.03-0.09g Lrax, 0.06-0.10g NAA, 600u PXB, 60 μ g amphotericin Bs, 240 μ g Nalidixic Acids, 60 μ g methoxy Bian amic metadiazines and 60 μ g azlocillins, deionized water 200ml.
Preferably, in said A liquid, said potassium primary phosphate is 1.5g, and said sal epsom is 0.307g, and said magnesium citrate is 0.584g, and said l-asparagine is 2.4g, and said agar powder is 15g; In said B liquid, said bovine serum albumin is 3.5g, and said glucose is 5g; Said katalase is 1.5g, and said oleic acid is 10g, and said Triphosaden is 0.06g; Said coenzyme A is 0.5g, and said Lrax is 0.05g, and said NAA is 0.08g.
Preferably, in said A liquid, said potassium primary phosphate is 2g, and said sal epsom is 0.5g, and said magnesium citrate is 0.5g, and said l-asparagine is 3g, and said agar powder is 20g; In said B liquid, said bovine serum albumin is 4g, and said glucose is 4.5g; Said katalase is 1.2g, and said oleic acid is 5g, and said Triphosaden is 0.10g; Said coenzyme A is 0.3g, and said Lrax is 0.03g, and said NAA is 0.10g.
Preferably, in said A liquid, said potassium primary phosphate is 1g, and said sal epsom is 0.3g, and said magnesium citrate is 0.8g, and said l-asparagine is 2g, and said agar powder is 18g; In said B liquid, said bovine serum albumin is 3g, and said glucose is 4g; Said katalase is 1.8g, and said oleic acid is 8g, and said Triphosaden is 0.01g; Said coenzyme A is 0.8g, and said Lrax is 0.09g, and said NAA is 0.06g.
Preferably, said A liquid also comprises the 0.02-0.03g ferric ammonium citrate.
Preferably, said l-asparagine substitutes with edible monosodium glutamate, and consumption doubles.
Preferably, said bovine serum albumin personnel selection or animal serum substitute.
Preferably, at least a using with the similar medicine of its antimicrobial spectrum replaces in said PXB, said amphotericin B, said Nalidixic Acid, said methoxy Bian amic metadiazine and the said azlocillin.
In second aspect of the present invention; A kind of standardized production method of above-mentioned fast-growing solid culture medium of tubercle bacillus is provided; Be characterized in; Potassium primary phosphate, said sal epsom, said magnesium citrate and said l-asparagine described in the said A liquid dissolve fully sterilizes after the back adds said agar mixing, is cooled to 45-50 ℃ naturally; Said B liquid dissolves the after-filtration degerming fully, adds rapidly mixing in the said A liquid after being incubated to 40-50 ℃.Generally packing as early as possible, bevel.
Preferably, the condition of said sterilization is 110 ℃ of high pressure 20 minutes, and the 0.22nm filtration sterilization is adopted in said filtration sterilization.
Beneficial effect of the present invention is specific as follows:
1. fast-growing solid culture medium of tubercle bacillus of the present invention with agar as propping material; Adding stimulates the mycobacterium tuberculosis nutritive substance of growth fast; Comprise and to accelerate the tubercule bacillus metabolic activity to improve energy matter such as ATP and the growth-promoting substance such as the px etc. of tubercule bacillus growth and breeding speed; Accelerate the speed of growth of tubercule bacillus greatly, designed ingenious, simple and convenient preparation, cost is low, steady quality is reliable (preservation period can reach more than 1 year), be easy to stdn scale operation;
2. contain basic substance in the fast-growing solid culture medium of tubercle bacillus of the present invention, comprise all components in the existing solid tubercule bacillus substratum in the basic substance, they are: potassium primary phosphate, sal epsom (MgSO
47H
2O), citron magnesium, asparagine, water, each component role is respectively: have the salts such as citrate of magnesia, sal epsom of shock absorption to supply with elements such as magnesium, potassium, the effect of regulating osmotic pressure is arranged; Make the cytolemma of tubercule bacillus that good permeability arranged, asparagine is a nitrogenous source, can promote growth and the breeding of tubercule bacillus; Potassium primary phosphate is a buffer reagent, regulates the medium pH value, and multiple microbiotic can suppress the growth of the assorted bacterium of part in the sample; In addition; Also have ferric ammonium citrate and bovine serum albumin etc. in the basic substance, ferric ammonium citrate can provide certain irony required to satisfy the tubercule bacillus growth and breeding, can increase bacterium as tubercule bacillus to cultivate usefulness; Therefore; Fast culture media for mycobaterium tuberculosis of the present invention is compared with existing solid tubercule bacillus substratum, and kind of nutritive substance is many in it, and the growth and breeding that it can be tubercule bacillus provides the sufficient nutrient thing;
3. fast-growing solid culture medium of tubercle bacillus of the present invention also contains energy matter and growth-promoting substance, and energy matter can quicken the metabolic activity of tubercule bacillus, can improve the growth and breeding speed of tubercule bacillus, and growth stimulator mass-energy promotes the growth of tubercule bacillus.
Embodiment
In order more to be expressly understood technology contents of the present invention, the special following examples of lifting specify.
Embodiment 1
A liquid: 1.5g potassium primary phosphate, 0.307g sal epsom, 0.584g magnesium citrate, 2.4g l-asparagine and 15g agar powder, 800ml deionized water;
B liquid: 3.5g bovine serum albumin, 5g glucose, 1.5g katalase, 10g oleic acid, 0.06g Triphosaden, 0.5g coenzyme A, 0.05g Lrax, 0.08g NAA, 600u PXB, 60 μ g amphotericin Bs, 240 μ g Nalidixic Acids, 60 μ g methoxy Bian amic metadiazines and 60 μ g azlocillins, deionized water 200ml.
Compound method: 110 ℃ of high pressure were sterilized in 20 minutes after potassium primary phosphate, said sal epsom, said magnesium citrate and said l-asparagine described in the said A liquid dissolved the said agar mixing of back adding fully, were cooled to 45-50 ℃ naturally; Said B liquid dissolves back 0.22nm filtration sterilization fully, adds mixing in the said A liquid rapidly after being incubated to 40-50 ℃.Packing is extremely in vitro a plurality of as early as possible, the bevel solid medium.
Different in vitro inoculating type strain H37Rv respectively, antitubercular agent complete responsive tuberculosis clinical strains (12 strain), drug-resistant type tuberculosis clinical strains (12 strain) being cultivated, find that all strains examined production is rapid, the L-J tuberculosis substratum of comparing traditional; Bacterium colony just forms and suitable judgement time greatly shifts to an earlier date; Just form 7-9 days, average 8.3 days, about in advance 3-4 days; Average 15.4 days of the suitable judgement time as a result of same colonial morphology, about in advance 10 days (about around the L-J substratum is general).
Embodiment 2
A liquid: 2g potassium primary phosphate, 0.5g sal epsom, 0.5g magnesium citrate, 3g l-asparagine and 20g agar powder, 800ml deionized water;
B liquid: 4g bovine serum albumin, 4.5g glucose, 1.2g katalase, 5g oleic acid, 0.10g Triphosaden, 0.3g coenzyme A, 0.03g Lrax, 0.10g NAA, 600u PXB, 60 μ g amphotericin Bs, 240 μ g Nalidixic Acids, 60 μ g methoxy Bian amic metadiazines and 60 μ g azlocillins, deionized water 200ml.
Compound method: with embodiment 1.
Different in vitro inoculating type strain H37Rv respectively, antitubercular agent complete responsive clinical strains (12 strain), drug-resistant type tuberculosis clinical strains (12 strain) are being cultivated; Find that all strains examined production is rapid; Grew respectively 15,16,18 days, size is with embodiment 1.
Embodiment 3
A liquid: 1g potassium primary phosphate, 0.3g sal epsom, 0.8g magnesium citrate, 2g l-asparagine and 18g agar powder, 800ml deionized water;
B liquid: 3g bovine serum albumin, 4g glucose, 1.8g katalase, 10g oleic acid, 0.01g Triphosaden, 0.8g coenzyme A, 0.09g Lrax, 0.06g NAA, 600u PXB, 60 μ g amphotericin Bs, 240 μ g Nalidixic Acids, 60 μ g methoxy Bian amic metadiazines and 60 μ g azlocillins, deionized water 200ml.
Compound method: with embodiment 1.
Different in vitro inoculating type strain H37Rv respectively, antitubercular agent complete responsive bacterial strain (12 strain), drug-resistant type tubercule bacillus (12 strain) being cultivated, find that all strains examined production is rapid, to give birth to 15,16,18 days respectively, size is with embodiment 1.
Embodiment 4
A liquid: 1g potassium primary phosphate, 0.3g sal epsom, 0.8g magnesium citrate, 2g l-asparagine, 0.02g ferric ammonium citrate and 18g agar powder, 800ml deionized water;
B liquid: 3g bovine serum albumin, 4g glucose, 1.8g katalase, 8g oleic acid, 0.01g Triphosaden, 0.8g coenzyme A, 0.09g Lrax, 0.06g NAA, 600u PXB, 60 μ g amphotericin Bs, 240 μ g Nalidixic Acids, 60 μ g methoxy Bian amic metadiazines and 60 μ g azlocillins, deionized water 200ml.
Compound method: 110 ℃ of high pressure were sterilized in 20 minutes after potassium primary phosphate, said sal epsom, said magnesium citrate, said l-asparagine and said ferric ammonium citrate described in the said A liquid dissolved the said agar mixing of back adding fully, were cooled to 45-50 ℃ naturally; Said B liquid dissolves back 0.22nm filtration sterilization fully, adds mixing in the said A liquid rapidly after being incubated to 40-50 ℃.Packing is extremely in vitro a plurality of as early as possible, the bevel solid medium.
Different in vitro inoculating type strain H37Rv respectively, antitubercular agent complete responsive bacterial strain (12 strain), drug-resistant type tubercule bacillus (12 strain) being cultivated, find that all strains examined production is rapid, to grow respectively 15,16,18 days, size is with embodiment 1.
Embodiment 5
A liquid: 1g potassium primary phosphate, 0.3g sal epsom, 0.8g magnesium citrate, 2g l-asparagine, 0.03g ferric ammonium citrate and 18g agar powder, 800ml deionized water;
B liquid: 3g bovine serum albumin, 4g glucose, 1.8g katalase, 10g oleic acid, 0.01g Triphosaden, 0.8g coenzyme A, 0.09g Lrax, 0.06g NAA, 600u PXB, 60 μ g amphotericin Bs, 240 μ g Nalidixic Acids, 60 μ g methoxy Bian amic metadiazines and 60 μ g azlocillins, deionized water 200ml.
Compound method: with embodiment 4.
Different in vitro inoculating type strain H37Rv respectively, antitubercular agent complete responsive bacterial strain (12 strain), drug-resistant type tubercule bacillus (12 strain) being cultivated, find that all strains examined production is rapid, to grow respectively 15,16,18 days, size is with embodiment 1.
Embodiment 6
A liquid: 2g potassium primary phosphate, 0.5g sal epsom, 0.5g magnesium citrate, the edible monosodium glutamate of 6g and 20g agar powder, 800ml deionized water;
B liquid: 4g bovine serum albumin, 4.5g glucose, 1.2g katalase, 5g oleic acid, 0.10g Triphosaden, 0.3g coenzyme A, 0.03g Lrax, 0.10g NAA, 600u PXB, 60 μ g amphotericin Bs, 240 μ g Nalidixic Acids, 60 μ g methoxy Bian amic metadiazines and 60 μ g azlocillins, deionized water 200ml.
Compound method: 110 ℃ of high pressure were sterilized in 20 minutes after potassium primary phosphate described in the said A liquid, said sal epsom, said magnesium citrate, said edible monosodium glutamate and said ferric ammonium citrate dissolved the said agar mixing of back adding fully, were cooled to 45-50 ℃ naturally; Said B liquid dissolves back 0.22nm filtration sterilization fully, adds mixing in the said A liquid rapidly after being incubated to 40-50 ℃.Packing is extremely in vitro a plurality of as early as possible, the bevel solid medium.。
Different in vitro inoculating type strain H37Rv respectively, antitubercular agent complete responsive bacterial strain (12 strain), drug-resistant type tubercule bacillus (12 strain) being cultivated, find that all strains examined production is rapid, to grow respectively 15,16,18 days, size is with embodiment 1.
Embodiment 7
A liquid: 2g potassium primary phosphate, 0.5g sal epsom, 0.5g magnesium citrate, 3g l-asparagine and 20g agar powder, 800ml deionized water;
B liquid: 4g human serum albumin, 4.5g glucose, 1.2g katalase, 5g oleic acid, 0.10g Triphosaden, 0.3g coenzyme A, 0.03g Lrax, 0.10g NAA, 600u PXB, 60 μ g amphotericin Bs, 240 μ g Nalidixic Acids, 60 μ g methoxy Bian amic metadiazines and 60 μ g azlocillins, deionized water 200ml.
Compound method: with embodiment 1.
Different in vitro inoculating type strain H37Rv respectively, antitubercular agent complete responsive bacterial strain (12 strain), drug-resistant type tubercule bacillus (12 strain) being cultivated, find that all strains examined production is rapid, to grow respectively 15,16,18 days, size is with embodiment 1.
Tubercule bacillus is that obligate is aerobic, and nutritional requirement is higher, poor growth; It is a facultative parasite in the born of the same parents, is a kind of unicellular organism, owing to have the hydrophobicity skin; Nutritive substance is difficult for absorbing, so poor growth, modified Russell medium is widespread use; But incubation time is oversize, can not satisfy the needs of clinical quick diagnosis and treatment.Comparing on the fast-growing solid culture medium of tubercle bacillus of the present invention more than the obvious shortening of growth required time can shift to an earlier date nearly 1 week on the modified Russell medium through the foregoing description discovery tubercule bacillus; The growth required time of different drug-resistant type tubercule bacilluss is also different; Than type strain H37Rv and longer to the complete responsive strain growth required time of antitubercular agent; Tubercule bacillus poor growth or general on modified Russell medium, and growth is all fine on novel fast-growing solid culture medium.Therefore, fast-growing solid culture medium of tubercle bacillus of the present invention can not only shorten tubercule bacillus growth required time, and makes tubercule bacillus ability dominant growth, improves positive separation rate, helps the clinical diagnosis of accurately making.Therefore, fast-growing solid culture medium of tubercle bacillus of the present invention can be used as Russell medium the improvement novel method in the widespread use of Clinical microorganism laboratory.
To sum up; Fast-growing solid culture medium of tubercle bacillus of the present invention design is ingenious, simple and convenient preparation, cost is low, steady quality is reliable, be easy to stdn scale operation; The tubercule bacillus growth of cultivating above that fast; Thereby make drug sensitive test fast, accurately, help the early diagnosis and therapy of tubercule bacillus.
In this specification sheets, the present invention is described with reference to its certain embodiments.But, still can make various modifications and conversion obviously and not deviate from the spirit and scope of the present invention.Therefore, specification sheets and accompanying drawing are regarded in an illustrative, rather than a restrictive.
Claims (10)
1. a fast-growing solid culture medium of tubercle bacillus is characterized in that, adopts A liquid and B liquid formulated with 1: 1 ratio, wherein:
A liquid: 1.0-2.0g potassium primary phosphate, 0.3-0.5g sal epsom, 0.5-0.8g magnesium citrate, 2.0-3.0g l-asparagine and 15-20g agar powder, 800ml deionized water;
B liquid: 3.0-4.0g bovine serum albumin, 4-5g glucose, 1.2-1.8g katalase, 5-10g oleic acid, 0.01-0.10g Triphosaden, 0.3-0.8g coenzyme A, 0.03-0.09g Lrax, 0.06-0.10g NAA, 600u PXB, 60 μ g amphotericin Bs, 240 μ g Nalidixic Acids, 60 μ g methoxy Bian amic metadiazines and 60 μ g azlocillins, deionized water 200ml;
Potassium primary phosphate, said sal epsom, said magnesium citrate and said l-asparagine described in the said A liquid dissolve fully sterilizes after the back adds said agar mixing, is cooled to 45-50 ℃ naturally; Said B liquid dissolves the after-filtration degerming fully, adds rapidly mixing in the said A liquid after being incubated to 40-50 ℃.
2. fast-growing solid culture medium of tubercle bacillus according to claim 1 is characterized in that, in said A liquid; Said potassium primary phosphate is 1.5g, and said sal epsom is 0.307g, and said magnesium citrate is 0.584g; Said l-asparagine is 2.4g, and said agar powder is 15g; In said B liquid, said bovine serum albumin is 3.5g, and said glucose is 5g; Said katalase is 1.5g, and said oleic acid is 10g, and said Triphosaden is 0.06g; Said coenzyme A is 0.5g, and said Lrax is 0.05g, and said NAA is 0.08g.
3. fast-growing solid culture medium of tubercle bacillus according to claim 1 is characterized in that, in said A liquid, said potassium primary phosphate is 2g, and said sal epsom is 0.5g, and said magnesium citrate is 0.5g, and said l-asparagine is 3g, and said agar powder is 20g; In said B liquid, said bovine serum albumin is 4g, and said glucose is 4.5g; Said katalase is 1.2g, and said oleic acid is 5g, and said Triphosaden is 0.10g; Said coenzyme A is 0.3g, and said Lrax is 0.03g, and said NAA is 0.10g.
4. fast-growing solid culture medium of tubercle bacillus according to claim 1 is characterized in that, in said A liquid, said potassium primary phosphate is 1g, and said sal epsom is 0.3g, and said magnesium citrate is 0.8g, and said l-asparagine is 2g, and said agar powder is 18g; In said B liquid, said bovine serum albumin is 3g, and said glucose is 4g; Said katalase is 1.8g, and said oleic acid is 8g, and said Triphosaden is 0.01g; Said coenzyme A is 0.8g, and said Lrax is 0.09g, and said NAA is 0.06g.
5. fast-growing solid culture medium of tubercle bacillus according to claim 1 is characterized in that, said A liquid also comprises the 0.02-0.03g ferric ammonium citrate.
6. fast-growing solid culture medium of tubercle bacillus according to claim 1 is characterized in that, said l-asparagine substitutes with edible monosodium glutamate, and consumption doubles.
7. fast-growing solid culture medium of tubercle bacillus according to claim 1 is characterized in that, said bovine serum albumin personnel selection or animal serum substitute.
8. fast-growing solid culture medium of tubercle bacillus according to claim 1; It is characterized in that at least a using with the similar medicine of its antimicrobial spectrum replaces in said PXB, said amphotericin B, said Nalidixic Acid, said methoxy Bian amic metadiazine and the said azlocillin.
9. the standardized production method of a fast-growing solid culture medium of tubercle bacillus according to claim 1; It is characterized in that; Potassium primary phosphate, said sal epsom, said magnesium citrate and said l-asparagine described in the said A liquid dissolve fully sterilizes after the back adds said agar mixing, is cooled to 45-50 ℃ naturally; Said B liquid dissolves the after-filtration degerming fully, adds rapidly mixing in the said A liquid after being incubated to 40-50 ℃.
10. standardized production method according to claim 9 is characterized in that, the condition of said sterilization is 110 ℃ of high pressure 20 minutes, and the 0.22nm filtration sterilization is adopted in said filtration sterilization.
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