BCYE Medium, Diphasic Blood Culture (Buffered Charcoal Yeast Extract Medium, Diphasic Blood Culture)

[所属分类:培养基配方] [发布时间:2019-12-10] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

BCYE Medium, Diphasic Blood Culture
(Buffered Charcoal Yeast Extract
Medium, Diphasic Blood Culture)

培养基配方
Composition per liter:
Agar phase ....................................................1.0L
Broth phase....................................................1.0L
pH 6.9 ± 0.2 at 25°C
Agar Phase:
Composition per liter:
Agar ..........................................................20.0g
ACES buffer (2-[(2-amino-2-oxoethyl)-
amino]-ethane sulfonic acid)....................................10.0g
Yeast extract...................................................10.0g
Charcoal, activated, acid washed................................4.0g
KOH.............................................................2.8g
α-Ketoglutarate................................................1.0g
L -Cysteine·HCl·H 2 O solution .............................10.0mL
Fe 4 (P 2 O 7 ) 3 ·9H 2 O solution...........................10.0mL
L -Cysteine·HCl·H 2 O Solution:
Composition per 10.0mL:
L- Cysteine·HCl·H 2 O........................................0.4g
Preparation of  L -Cysteine·HCl·H2O Solution: Add  L -
cysteine·HCl·H2O to distilled/deionized water and bring volume to
10.0mL. Mix thoroughly. Filter sterilize.
Fe4(P2O7)3 ·9H2O Solution:
Composition per 10.0mL:
Fe4(P2O7)3 ·9H2O..............................................0.25g
Preparation of Fe4(P2O7)3 ·9H2O Solution: Add Fe4 (P2O7)3 ·9H2O
to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Filter sterilize.
Preparation of Agar Phase:  Add components, except  L -
cysteine·HCl·H2O solution and Fe4(P2O7)3 solution, to distilled/deion-
ized water and bring volume to 980.0mL. Mix thoroughly. Adjust me-
dium to pH 6.9 with 1N KOH. Heat gently and bring to boiling for 1
min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50°–
55°C. Aseptically add the  L -cysteine·HCl·H2O solution and
Fe 4 (P2O7)3 ·9H2O solution. Mix thoroughly.
Broth Phase:
Composition per liter:
ACES buffer (2-[(2-amino-2-oxoethyl)-
amino]-ethane sulfonic acid)...............................10.0g
Yeast extract..............................................10.0g
Charcoal, activated, acid washed............................4.0g
KOH.........................................................2.4g
α-Ketoglutarate............................................1.0g
Sodium polyaneolsulfonate .................................0.3g
L -Cysteine·HCl·H 2 O solution..........................10.0mL
Fe4(P2O7)3 ·9H2O solution................................10.0mL
L -Cysteine·HCl·H2O Solution:
Composition per 10.0mL:
L- Cysteine·HCl·H2O......................................0.4g
Preparation of  L -Cysteine·HCl·H2O Solution: Add  L -
cysteine·HCl·H2O to distilled/deionized water and bring volume to
10.0mL. Mix thoroughly. Filter sterilize.
Fe4(P2O7)3 ·9H2O Solution:
Composition per 10.0mL:
Fe4(P2O7)3 ·9H2O..............................................0.25g
Preparation of Fe4(P2O7)3 ·9H2O Solution: Add Fe4(P207)3 ·9H2O
to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Filter sterilize.
Preparation of Broth Phase:  Add components, except  L -
cysteine·HCl·H2O solution and Fe4(P2O7)3 solution, to distilled/deion-
ized water and bring volume to 980.0mL. Mix thoroughly. Adjust me-
dium to pH 6.9 with 1N KOH. Heat gently and bring to boiling for 1
min. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 50–55°C.
Aseptically add the cysteine·HCl·H2O solution and Fe4(P2O7)3 ·9H2O
solution. Mix thoroughly.
Preparation of Medium: Aseptically distribute cooled sterile agar
phase into sterile blood culture bottles in 100.0mL volumes. Allow
bottles to cool in a slanted position. Aseptically add 50.0mL of sterile
broth phase to each blood culture bottle.
Use: For the isolation and cultivation of Legionella pneumophila and
other Legionella species from blood samples.