FRAG Agar

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

FRAG Agar (Fragilis Agar) 

培养基配方 

Composition per 1025.0mL:
L- Cysteine·HCl·H 2 O......................................................................0.5g
Basal solution.........................................................................995.0mL
Glucuronic acid solution..........................................................25.0mL
Gentamicin solution...................................................................1.0mL
Hemin-vitamin K 1 solution........................................................1.0mL
Ferric sulfate solution ................................................................1.0mL
Mineral solution.........................................................................1.0mL
Phenol Red (1% solution)..........................................................1.0mL
Vitamin B 12 solution................................................................0.05mL
pH 7.0 ± 0.1 at 25°C
Basal Solution:
Composition per 995.0mL:
Oxgall..........................................................................................20.0g
Agar ............................................................................................15.4g
K 2 HPO 4 .......................................................................................2.26g
Yeast extract..................................................................................2.0g
Pancreatic digest of casein............................................................1.4g
(NH 4 ) 2 HPO 4 ..................................................................................1.0g
K 2 HPO 4 .........................................................................................0.9g
Papaic digest of soybean meal....................................................0.12g
NaCl............................................................................................0.12g
Preparation of Basal Solution: Add components to distilled/deionized water and bring volume to 995.0mL. Mix thoroughly. Gently heat and bring to boiling. Autoclave for 15 min at 15 psi pressure– 121°C. Cool to 45°–50°C.
Glucuronic Acid Solution:
Composition per 100.0mL:
D- Glucuronic acid........................................................................40.0g
Preparation of Glucuronic Acid Solution: Add glucuronic acid to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.

Gentamicin Solution:
Composition per 10.0mL:
Gentamicin.................................................................................0.1mg
Preparation of Gentamicin Solution: Add gentamicin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Vitamin K 1 -Hemin Solution:
Composition per liter:
Vitamin K 1 solution.................................................................10.0mL
Hemin solution.........................................................................10.0mL
Preparation of Vitamin K 1 -Hemin Solution: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Filter sterilize.
Vitamin K 1 Solution:
Composition per 100.0mL:
Vitamin K 1 ....................................................................................1.0g
Ethanol.....................................................................................99.0mL
Preparation of Vitamin K 1 Solution: Add vitamin K 1 to 99.0mL of absolute ethanol. Mix thoroughly.
Hemin Solution:
Composition per 10.0mL:
Hemin ...........................................................................................0.5g
NaOH............................................................................................0.4g
Preparation of Hemin Solution:  Add hemin and NaOH to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Ferric Sulfate Solution:
Composition per 100.0mL:
FeSO 4 ·9H 2 O................................................................................0.04g
Preparation of Ferric Sulfate Solution:  Add FeSO 4 ·9H 2 O to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Mineral Solution:
Composition per 100.0mL:
NaCl..............................................................................................9.0g
CaCl 2 ·2H 2 O ................................................................................0.27g
MgCl 2 ·6H 2 O .................................................................................0.2g
CoCl 2 ·6H 2 O..................................................................................0.1g
MnCl 2 ·4H 2 O .................................................................................0.1g
Preparation of Mineral Solution:  Add components to distilled/ deionized water and bring volume to 100.0mL. Mix thoroughly. Filter sterilize.
Vitamin B 12 Solution:
Composition per 10.0mL:
Vitamin B 12 ................................................................................0.1mg
Preparation of Vitamin B 12 Solution: Add vitamin B 12 to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: To 995.0mL of cooled, sterile basal solution, aseptically add 0.5g of  L -cysteine·HCl·H 2 O, 25.0mL of sterile glucuronic acid solution, 1.0mL of sterile gentamicin solution, 1.0mL of sterile hemin-vitamin K 1 solution, 1.0mL of sterile ferric sulfate solution, 1.0mL of sterile mineral solution, 1.0mL of Phenol Red solution, and 0.05mL of sterile vitamin B 12 solution. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.
Use: For the isolation, cultivation, and differentiation of the Bacteroides fragilis group (Bacteroides fragilis, Bacteroides thetaiotamicron, Bacteroides vulgatus, Bacteroides distasonis, Bacteriodes ovatus, and Bacteroides uniformis) from clinical specimens.