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Fast-growing solid culture medium of tubercle bacillus and standardized production method thereof

A solid medium and tuberculosis technology, applied in tuberculosis culture and biological fields, can solve the problems of slow growth of tubercle bacilli and the inability to provide guidance for clinical rational drug use, etc., and achieve easy standardized large-scale production, accelerated metabolic activities, and stable quality reliable effect

Inactive Publication Date: 2010-01-27
无锡博慧斯生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the slow growth of Mycobacterium tuberculosis, this traditional method usually takes 4-6 weeks to report the results of the drug susceptibility test, so it cannot provide timely guidance for clinical rational drug use

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Solution A: 1.5g potassium dihydrogen phosphate, 0.307g magnesium sulfate, 0.584g magnesium citrate, 2.4g asparagine, 15g agar powder, 800ml deionized water;

[0030] Solution B: 3.5g bovine serum albumin, 5g glucose, 1.5g catalase, 10g oleic acid, 0.06g adenosine triphosphate, 0.5g coenzyme A, 0.05g cytochrome C, 0.08g α-naphthaleneacetic acid, 600u polymyxin B. 60 μg amphotericin B, 240 μg nalidixic acid, 60 μg trimethoprim and 60 μg azlocillin, 200 ml deionized water.

[0031]Preparation method: The potassium dihydrogen phosphate, the magnesium sulfate, the magnesium citrate, and the asparagine in the liquid A are completely dissolved, then added to the agar and mixed evenly, and then sterilized under high pressure at 110°C for 20 minutes. Lower the temperature to 45-50°C; after the liquid B is completely dissolved, sterilize it by 0.22nm filtration, and after incubating to 40-50°C, quickly add it to the liquid A and mix well. Divide into multiple test tubes as soon...

Embodiment 2

[0034] Solution A: 2g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.5g magnesium citrate, 3g asparagine, 20g agar powder, 800ml deionized water;

[0035] Solution B: 4g bovine serum albumin, 4.5g glucose, 1.2g catalase, 5g oleic acid, 0.10g adenosine triphosphate, 0.3g coenzyme A, 0.03g cytochrome C, 0.10g α-naphthaleneacetic acid, 600u polymyxin B. 60 μg amphotericin B, 240 μg nalidixic acid, 60 μg trimethoprim and 60 μg azlocillin, 200 ml deionized water.

[0036] Preparation method: with embodiment 1.

[0037] The standard strain H37Rv, the clinical strains (12 strains) which are all sensitive to anti-tuberculosis drugs, and the clinical strains (12 strains) of drug-resistant tuberculosis were inoculated in different test tubes, and it was found that all the strains produced rapidly and grew 15, 16, and 18 respectively. Day, size is the same as embodiment 1.

Embodiment 3

[0039] Solution A: 1g potassium dihydrogen phosphate, 0.3g magnesium sulfate, 0.8g magnesium citrate, 2g asparagine, 18g agar powder, 800ml deionized water;

[0040] Solution B: 3g bovine serum albumin, 4g glucose, 1.8g catalase, 10g oleic acid, 0.01g adenosine triphosphate, 0.8g coenzyme A, 0.09g cytochrome C, 0.06g α-naphthaleneacetic acid, 600u polymyxin B , 60 μg amphotericin B, 240 μg nalidixic acid, 60 μg trimethoprim and 60 μg azlocillin, 200 ml deionized water.

[0041] Preparation method: with embodiment 1.

[0042] Inoculate standard strain H37Rv, anti-tuberculosis drug all-sensitive strains (12 strains), and drug-resistant tuberculosis bacteria (12 strains) in different test tubes for culture, and found that all strains produced rapidly, and they were born for 15, 16, and 18 days respectively. Size is with embodiment 1.

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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 [0001] The invention relates to the technical field of biology, in particular to the technical field of Mycobacterium tuberculosis culture, in particular to a solid culture medium for rapid growth of Mycobacterium tuberculosis and a standardized production method thereof. Background technique [0002] For a long time, the laboratory diagnosis of tuberculosis has mainly relied on bacteriological smear and culture examination. Due to the limitations of the method, it has seriously affected the timely diagnosis of tuberculosis. Therefore, domestic and foreign scholars have been looking for rapid, sensitive, specific and simple laboratory diagnostic methods for many years. [0003] 1. Smear and culture [0004] Smear and culture are the most basic bacteriological examination methods for tuberculosis. The sensitivity of culture is slightly higher than that of smear method, and it is a reliable method to identify live bacteria. It can also be further used for ba...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/32
Inventor 孙战强张舒林雷震
Owner 无锡博慧斯生物医药科技有限公司
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