Haemophilus ducreyi Medium, Revised (Ducreyi Medium, Revised)

[所属分类:培养基配方] [发布时间:2021-3-23] [发布人: ] [阅读次数:] [返回]

Haemophilus ducreyi Medium, Revised

(Ducreyi Medium, Revised)

培养基配方

Composition per 1010.0mL:
Solution B..............................................................................500.0mL
Solution A..............................................................................400.0mL
Solution C..............................................................................110.0mL
pH 7.4 ± 0.2 at 25°C
Solution A:
Composition per 400.0mL:
Beef heart, infusion from..........................................................500.0g
Agar............................................................................................15.0g
Tryptose......................................................................................10.0g
NaCl..............................................................................................5.0g
Preparation of Solution A: Add components to distilled/deionized water and bring volume to 400.0L. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C.
Solution B:
Composition per 500.0mL:
Hemoglobin................................................................................10.0g
Preparation of Solution B: Add hemoglobin to distilled/deionized water and bring volume to 500.0L. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C.
Solution C:
Composition per 110.0mL:
Fetal bovine serum.................................................................100.0mL
Supplement solution................................................................10.0mL
Supplement Solution:
Composition per liter:
Glucose.....................................................................................100.0g
L -Cysteine·HCl ...........................................................................25.9g
L -Glutamine................................................................................10.0g
L -Cystine.......................................................................................1.1g
Adenine.........................................................................................1.0g
Nicotinamide adenine dinucleotide ............................................0.25g
Vitamin B 12 ...................................................................................0.1g
Thiamine pyrophosphate ..............................................................0.1g
Guanine·HCl...............................................................................0.03g
Fe(NO 3 ) 3 ·6H 2 O...........................................................................0.02g
p-Aminobenzoic acid................................................................0.013g
Thiamine·HCl ............................................................................3.0mg
Source: The supplement solution (IsoVitaleX ® enrichment) is available from BD Diagnostic Systems. This enrichment may be replaced by supplement VX from BD Diagnostic Systems.
Preparation of Supplement Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.
Preparation of Solution C: Combine components. Mix thoroughly. Filter sterilize. Warm to 45°–50°C. 

Preparation of Medium: Aseptically combine solution A, solution B, and solution C. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.

Use:  For the cultivation and maintenance of Haemophilus ducreyi.