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Bartonia body solid-liquid double-phase slant culture medium as well as preparation method and application method thereof

A technology of slant culture medium and solid culture medium, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of inability to study bacterial secreted proteins, long growth cycle and high cost

Inactive Publication Date: 2010-09-22
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] (1) The growth cycle is longer;
[0005] (2) Large-scale cultivation requires a large number of plates, which consumes a lot of manpower, material resources and time;
[0006] (3) Blood plate culture can only collect colonies, but cannot study bacterial secreted proteins
However, the formula of the liquid medium is complex, requiring more than 30 substances such as NAD, NADP, various amino acid salts, etc., the cost is high, and the requirements for reagent configuration and experimenters are also strict.

Method used

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  • Bartonia body solid-liquid double-phase slant culture medium as well as preparation method and application method thereof
  • Bartonia body solid-liquid double-phase slant culture medium as well as preparation method and application method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Embodiment 1 Bartonella henselae (Bartonella henselae) solid-liquid two-phase medium configuration and cultivation

[0025] (1) Blood agar solid slant medium and blood agar plate medium

[0026] Beef extract 23.0g, starch 1.0g, sodium chloride 5.0g, agar 12.0g, water 1L, heated to 50°C to promote its dissolution. Pack into appropriate Erlenmeyer flasks and autoclave at 121°C for 15 minutes.

[0027] Preparation of defibrinated sheep blood: Collect 100ml of venous blood from healthy sheep, pour it into a sterile beaker, add 20 sterilized glass beads, shake at 120rpm / min for 10min, discard the glass beads, and store the blood at 4°C until use.

[0028] After the culture medium was cooled to 50°C, a total amount of 5% defibrinated sheep blood was added, and 3ml of each tube was poured into a sterilized test tube and placed obliquely to make a blood agar slant medium.

[0029] (2) Casein Trypsin Hydrolyzed Soybean Liquid Medium

[0030] Casein trypsin hydrolyzate 17g, so...

Embodiment 2

[0036] Embodiment 2 Bacillus-like Bartonella (Bartonella bacilliformis) solid-liquid biphasic medium configuration and cultivation

[0037] (1) Blood agar solid slant medium and blood agar plate medium

[0038] 30.0g of beef extract, 1.0g of starch, 5.0g of sodium chloride, 12.0g of agar, and 1000ml of water are heated to promote their dissolution. Pack into appropriate Erlenmeyer flasks and autoclave at 121°C for 15 minutes.

[0039] Preparation of defibrinated sheep blood: Collect 100ml of venous blood from healthy sheep, pour it into a sterile beaker, add 20 sterilized glass beads, shake at 120rpm / min for 10min, discard the glass beads, and store the blood at 4°C until use.

[0040] After the culture medium was cooled to 50°C, a total amount of 8% defibrinated sheep blood was added, and 3ml of each tube was poured into a sterilized test tube and placed obliquely to make a blood agar slant medium.

[0041] (2) Casein Trypsin Hydrolyzed Soybean Liquid Medium

[0042] Casei...

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Abstract

The invention relates to a bartonia body solid-liquid double-phase slant culture medium as well as a preparation method and an application method thereof. The slant culture medium comprises a slant solid culture medium and a liquid culture medium, wherein the slant solid culture medium comprises beef leach liquid, starch, sodium chloride, agar, defibrinated sheep blood and distilled water; and the liquid culture medium comprises trypticase hydrolysate, soybean protein and papayotin hydrolysate, glucose, sodium chloride, di-potassium phosphate and distilled water. The bartonia body solid-liquid double-phase slant culture medium prepared by the method approximately requires 48 hours to reach 107 CFU / ml, the growth speed is increased by 2.5 times, the cost is saved by 500 times (blood plate culture approximately requires 5 days, and reaching 107 CFU / ml requires 100 flat plates), and the defects of slow bacteria growth, time taking, labor taking, high cost and the like of the traditional bartonia body invitro culture method are remedied .

Description

technical field [0001] The invention relates to a culture medium and a preparation method in the technical field of biology, in particular to a Bartonella solid-liquid two-phase slant culture medium and a preparation and application method thereof. Background technique [0002] Bartonella is a group of Gram-negative, oxidase-negative, demanding nutritional conditions and facultative intracellular parasitic aerobic bacteria. Bartonella has included 19 species and subspecies, of which 7 are known to cause human diseases , which are bacillus-like, Hansai, five-day fever, Elizabeth, Graham, Klebsiella and Bartonella wenchenii Berghoff subspecies Bartonella. Among them, Bartonella henselae is the Bartonella species with the widest pathogenic spectrum. It can not only cause bacillary hemangioma-bacillary purpura in immunodeficiency patients, but also cause human cat scratch disease, chronic lymphadenopathy, endocarditis, optic nerve diseases such as inflammation. [0003] Barton...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
Inventor 袁聪俐华修国杨志彪崔立朱建国杨筱薇
Owner SHANGHAI JIAO TONG UNIV
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