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Diphasic quick differential medium of mycobacterium tuberculosis and application of medium

A technology for the identification of Mycobacterium tuberculosis culture medium, applied in the direction of microorganism-based methods, measurement/inspection of microorganisms, microorganisms, etc., can solve the problems of long cultivation and identification time, long vaccine production time, etc., and achieve growth and cultivation speed , The effect of shortening the cultivation time and preventing the contamination of bacteria

Inactive Publication Date: 2014-08-20
JINING MEDICAL UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

The invention is used to solve the problems of long time for culture and identification of Mycobacterium tuberculosis and long time for vaccine production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0043] (1) Preparation of solid medium:

[0044] a. Weigh 1~5mg of pyridoxine hydrochloride, 180~220mg of sodium glutamate, 10~20mg of calcium chloride, 15~25mg of magnesium sulfate, 80~120mg of ferric ammonium citrate, 180~220mg of sodium citrate, ammonium sulfate 380~420mg, sodium pyruvate 950~1050mg, copper chloride 1~10mg, zinc chloride 1~10mg, asparagine 1950~2050mg, potassium dihydrogen phosphate 15~25mg and sodium dihydrogen phosphate 15~25mg dissolved in distilled water In 600-700ml, heat at 121°C for 15-20 minutes, cool to 20-25°C to obtain solid medium base solution;

[0045] b. Weigh 8ml of neutral glycerin, 15ml of 1% malachite green aqueous solution and 1000ml of chicken egg liquid, add it to the solid medium base solution obtained in step a, mix evenly, and divide into sterile culture tubes, 7-10ml per tube;

[0046] c. Heat the sterile culture tube obtained in step b at 80-90°C for 10-20 minutes, cool to 20-25°C to obtain a solid medium, and store the solid cul...

Embodiment 1

[0073] (1) Preparation of solid medium:

[0074] a. Weigh pyridoxine hydrochloride 4mg, sodium glutamate 180mg, calcium chloride 15mg, magnesium sulfate 25mg, ferric ammonium citrate 90mg, sodium citrate 200mg, ammonium sulfate 380mg, sodium pyruvate 950mg, cupric chloride 5mg, Dissolve 5 mg of zinc chloride, 2000 mg of asparagine, 20 mg of potassium dihydrogen phosphate and 25 mg of sodium dihydrogen phosphate in 700 ml of distilled water, heat at 121°C for 20 minutes, and cool to 21°C to obtain a solid medium base solution;

[0075] b. Weigh 8ml of neutral glycerin, 15ml of 1% malachite green aqueous solution and 1000ml of chicken egg liquid, add it to the solid medium base solution obtained in step a, mix evenly, and divide into sterile culture tubes, 10ml per tube;

[0076] c. Heat the sterile culture tube obtained in step b at 90°C for 10 minutes, cool to 20-25°C to obtain a solid medium, and store the solid culture at 4°C for later use;

[0077] (2) Preparation of liquid ...

Embodiment 2

[0086] Preparation of liquid medium:

[0087] a. Weigh 5 mg of calcium chloride, 10 mg of magnesium sulfate, 18 mg of oleic acid, 10 mg of ferric ammonium citrate, 50 mg of sodium citrate, 200 mg of ammonium sulfate, 5 mg of copper chloride, 10 mg of zinc chloride, 450 mg of sodium pyruvate, and Wen-80 0.75ml, asparagine 1000mg, potassium dihydrogen phosphate 1000mg, sodium dihydrogen phosphate 1800mg were dissolved in 500ml of distilled water, heated at 121°C for 20 minutes, cooled to 22°C to obtain liquid medium base solution;

[0088] b. Weigh 0.15g of pyridoxine hydrochloride, 5ml of glycerin, 4ml of yeast extract, 0.2g of catalase, 0.15g of biotin, 0.6g of polyoxyethylene stearate, 200g of bovine serum albumin, and 200g of casein digestion Add 2g of the product and 5ml of the mycobacterium culture supernatant to 500ml of potato juice, mix well, filter and sterilize to obtain the growth-promoting factor mixed solution;

[0089] c. Take 500ml of liquid medium base solution...

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Abstract

The invention relates to a preparation method of a diphasic quick differential medium of mycobacterium tuberculosis, a method for quickly differentiating the mycobacterium tuberculosis by using the medium, a method for testing the drug resistance on antituberculous drugs caused by the mycobacterium tuberculosis and a method for quickly culturing a bacillus Calmette and Guerin vaccine by using a liquid medium in the medium. The diphasic medium prepared by adopting the method can be used for quickly culturing the mycobacterium tuberculosis, the diphasic quick culture medium is capable of differentiating the mycobacterium tuberculosis within 2-5 days, the differentiated mycobacterium tuberculosis can be subjected to drug resistance detection of the antituberculous drugs such as rifampicin, isoniazide and streptomycin by virtue of a reverse transcription PCR (polymerase chain reaction) method, and the liquid medium prepared by adopting the method is capable of finishing the culture of the bacillus Calmette and Guerin vaccine within 6-9 days.

Description

technical field [0001] The invention relates to the fields of bacterial culture and vaccine preparation. It specifically relates to a preparation method of a two-phase rapid identification medium for Mycobacterium tuberculosis, a method for rapidly identifying Mycobacterium tuberculosis using the medium, a method for measuring the drug resistance of Mycobacterium tuberculosis to anti-tuberculosis drugs, and a method for using the medium. A method for quickly cultivating BCG in liquid medium. Background technique [0002] Mycobacterium tuberculosis, commonly known as Mycobacterium tuberculosis, is the pathogenic bacteria that causes tuberculosis. It can invade all organs of the body, but tuberculosis is the most common. Tuberculosis is still an important infectious disease. An estimated one-third of the world's population is infected with Mycobacterium tuberculosis. According to WHO reports, about 1 out of every 3 people in the world is infected with Mycobacterium tubercu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12Q1/68C12R1/32
Inventor 薛庆节闫迎春聂尚丹李秀真章洪华杨媛媛胡文洁
Owner JINING MEDICAL UNIV
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