Legionella pneumophila Medium (Charcoal Yeast Extract Diphasic Blood Culture Medium) (Diphasic Blood Culture Buffered Charcoal Yeast Extract Medium) (CYE-DBCM)

[所属分类:培养基配方] [发布时间:2021-4-30] [发布人:李佳颐] [阅读次数:] [返回]

Legionella pneumophila Medium
(Charcoal Yeast Extract Diphasic
Blood Culture Medium)
(Diphasic Blood Culture Buffered
Charcoal Yeast Extract Medium)
(CYE-DBCM)

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

Agar phase .............................................................................500.0mL
Broth phase............................................................................500.0mL

pH 6.9–7.0 at 25°C

Agar Phase:
Composition per 500.0mL:
Agar............................................................................................17.0g
Charcoal, activated .......................................................................2.0g
Preparation of Agar Phase: Add components to distilled/deion-
ized water and bring volume to 500.0mL. Mix thoroughly. Gently heat
and bring to boiling. Distribute in 20.0mL volumes into 125.0mL se-
rum bottles with aluminum crimp seals and rubber stoppers. Autoclave
for 20 min at 15 psi pressure–121°C. Cool to 50°C. Swirl medium to
put charcoal in suspension. Allow agar to solidify so that a slant with a
6.0cm height is formed.
Broth Phase:
Composition per 500.0mL:
Yeast extract................................................................................20.0g
L- Cysteine·HCl·H 2 O solution........................................................0.4g
Fe(NO 3 ) 3 ·9H 2 O solution...............................................................0.1g
Preparation of Broth Phase: Add yeast extract to distilled/deion-
ized water and bring volume to 480.0mL. Mix thoroughly. Autoclave
for 15 min at 15 psi pressure–121°C. Cool to 25°C. Aseptically add
sterile  L -cysteine·HCl·H 2 O solution and Fe(NO 3 ) 3 ·9H 2 O solution. Mix
thoroughly. Adjust pH to 6.9 with 6.0mL of sterile 1N KOH.
L- Cysteine·HCl·H 2 O Solution:
Composition per 10.0mL:
L - Cysteine·HCl·H 2 O ...................................................................0.04g
Preparation of  L- Cysteine·HCl·H 2 O Solution: Add  L -cyste-
ine·HCl·H 2 O to distilled/deionized water and bring volume to 10.0mL.
Mix thoroughly. Filter sterilize.
Fe(NO 3 ) 3 ·9H 2 O Solution:
Composition per 10.0mL:
Fe(NO 3 ) 3 ·9H 2 O...........................................................................0.04g
Preparation of Fe(NO 3 ) 3 ·9H 2 O Solution: Add Fe(NO 3 ) 3 ·9H 2 O
to distilled/deionized water and bring volume to 10.0mL. Mix thor-
oughly. Filter sterilize.
Preparation of Medium: Add 20.0mL of sterile broth phase to
125.0mL serum bottles containing 20.0mL of solidified agar phase.
Seal bottles by crimping metal caps over rubber stoppers.
Use: For the isolation and cultivation of Legionella pneumophila from
blood cultures.