HBT Bilayer Medium (Human Blood Tween™ Bilayer Medium)

[所属分类:培养基配方] [发布时间:2021-3-31] [发布人: ] [阅读次数:] [返回]

HBT Bilayer Medium

(Human Blood Tween™ Bilayer Medium)

培养基配方

Composition per 1062.5mL:
Agar ............................................................................................13.5g
Pancreatic digest of casein..........................................................12.0g
Casein/meat peptone...................................................................10.0g
NaCl..............................................................................................5.0g
Peptic digest of animal tissue........................................................5.0g
Beef extract...................................................................................3.0g
Yeast extract..................................................................................3.0g
Cornstarch.....................................................................................1.0g
Human blood, anticoagulated..................................................25.0mL
Colistin solution.......................................................................10.0mL
Nalidixic acid solution.............................................................10.0mL
Amphotericin B solution..........................................................10.0mL
Polysorbate 80 (Tween™ 80) solution.......................................7.5mL
pH 7.3 ± 0.2 at 25°C
Source:  This medium is available as a premixed powder from BD Diagnostic Systems.
Colistin Solution:
Composition per liter:
Colistin........................................................................................0.01g
Preparation of Colistin: Add colistin to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Nalidixic Acid Solution:
Composition per liter:
Nalidixic acid..............................................................................0.02g
Preparation of Nalidixic Acid Solution: Add nalidixic acid to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Amphotericin B Solution:
Composition per liter:
Amphotericin B..........................................................................3.0mg
Preparation of Amphotericin B Solution: Add Amphotericin Bto distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Tween™ 80 Solution:
Composition per 100.0mL:
Tween™ 80................................................................................1.0mL
Preparation of Tween™ 80 Solution: Add Tween™ 80 to distilled/deionized water and bring volume to 100.0mL. Mix thoroughly. Adjust pH to 7.3. Filter sterilize.
Preparation of Medium: Add components, except amphotericin B solution, Tween™ 80, and human blood, to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Divide the medium into two 500.0mL fractions. Autoclave both flasks of media for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C. To one flask, aseptically add 5.0mL of sterile colistin solution, 5.0mL of sterile nalidixic acid solution, 5.0mL of sterile amphotericin B solution, and 3.75mL of Tween™ 80 solution. Mix thoroughly. Pour into sterile Petri dishes in 7.0mL volumes. Allow agar to harden. To remaining flask aseptically add 5.0mL of sterile colistin solution, 5.0mL of sterile nalidixic acid solution, 5.0mL of sterile amphotericin B solution, 3.75mL of sterile Tween™ 80 solution, and 25.0mL of sterile human blood. Mix thoroughly. Pour into the same Petri dishes that each contain 7.0mL of the agar medium without blood. The top layer should be approximately 14.0mL per plate.
Use: For the selective isolation, cultivation, and differentiation of Gardnerella vaginalis from clinical specimens.