GC Agar with Supplement A and VCTN Inhibitor

[所属分类:培养基配方] [发布时间:2021-3-9] [发布人: ] [阅读次数:] [返回]

GC Agar with Supplement A and VCTN Inhibitor

培养基配方

Composition per 1030.0mL:
GC medium base, 2X.............................................................500.0mL
Hemoglobin solution .............................................................500.0mL
Supplement solution ................................................................10.0mL
Supplement A..........................................................................10.0mL
VCTN inhibitor........................................................................10.0mL
pH 7.2 ± 0.2 at 25°C
GC Medium Base, 2X:
Composition per 500.0mL:
Proteose peptone No. 3...............................................................15.0g
Agar............................................................................................10.0g
NaCl..............................................................................................5.0g
K 2 HPO 4 .........................................................................................4.0g
Cornstarch.....................................................................................1.0g
KH 2 PO 4 .........................................................................................1.0g
Source:  GC medium base and Supplement A are available as a premixed powder from BD Diagnostic Systems.
Preparation of GC Medium Base, 2X: Add components to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Gently heat until boiling. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°–50°C.
Hemoglobin Solution:
Composition per 500.0mL:
Bovine hemoglobin.....................................................................10.0g
Preparation of Hemoglobin Solution: Add bovine hemoglobin to distilled/deionized water and bring volume to 500.0mL. Mix thoroughly. Autoclave for 15 min at 15 psi pressure–121°C. Cool to 45°– 50°C.
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.
Supplement A:
Composition per 10.0mL:
Supplement A contains yeast concentrate with Crystal Violet. Preparation of Supplement A:  Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
VCTN Inhibitor:
Composition per liter:
Colistin.......................................................................................7.5mg
Trimethoprim lactate..................................................................5.0mg
Vancomycin ...............................................................................4.0mg
Nystatin..................................................................................12,500U
Preparation of VCTN Inhibitor: Add components to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium: To 500.0mL of sterile GC medium base, aseptically add 500.0mL of sterile hemoglobin solution at 45°–50°C. Mix thoroughly. Aseptically add 10.0mL of sterile supplement solution, 10.0mL of sterile supplement A, and 10.0mL of sterile VCTN inhibitor. Mix thoroughly. Pour into sterile Petri dishes or distribute into sterile tubes.
Use: For the cultivation of Neisseria gonorrhoeae, other Neisseria
species, and Haemophilus species.