K-L Virulence Agar (Klebs-Loeffler Virulence Agar)

[所属分类:培养基配方] [发布时间:2021-4-23] [发布人:李佳颐] [阅读次数:] [返回]

K-L Virulence Agar
(Klebs-Loeffler Virulence Agar)

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

Composition per 1250.0mL:

K-L agar base................................................................................1.0L
K-L enrichment......................................................................200.0mL
K 2 TeO 3 solution.......................................................................50.0mL
K-L filter strips..............................................................................100
pH 7.8 ± 0.2 at 25°C
Source:  This medium is available as a premixed powder from BD Di-
agnostic Systems.
K-L Agar Base:
Composition per liter:
Meat peptone ..............................................................................20.0g
Agar............................................................................................15.0g
NaCl..............................................................................................2.5g
Preparation of K-L Agar Base: Add components to distilled/de-
ionized water and bring volume to 1.0L. Mix thoroughly. Gently heat
and bring to boiling. Autoclave for 15 min at 15 psi pressure–121°C.
Cool to 50°C.
K-L Enrichment:
Composition per 200.0mL:
Casamino acids.............................................................................4.0g
Glycerol .................................................................................100.0mL
Tween™ 80............................................................................100.0mL
Preparation of K-L Enrichment: Combine components. Mix
thoroughly. Filter sterilize.
K 2 TeO 3 Solution:
Composition per 100.0mL:
K 2 TeO 3 ..........................................................................................1.0g
Preparation of K 2 TeO 3 Solution:  Add K 2 TeO 3 to distilled/deion-
ized water and bring volume to 100.0mL. Mix thoroughly. Filter ster-
ilize.
Caution: Potassium tellurite is toxic.
K-L Filter Strips:
Composition :
Diphtheria toxin solution.........................................................10.0mL
Whatman No. 3 filter paper ................................................. as needed
Preparation of K-L Strips:  Cut Whatman #3 filter paper into
1.5cm × 7cm strips. Autoclave for 15 min at 15 psi pressure–121°C.
Aseptically dip each strip into a sterile solution containing 1000U of
purified diphtheria toxin/mL. Drain off excess liquid.
Preparation of Medium: To 1.0L of cooled, sterile K-L agar base,
aseptically add sterile K-L enrichment and sterile K 2 TeO 3 solution.
Mix thoroughly. Pour into sterile Petri dishes in 13.0mL volumes. Be-
fore the agar solidifies, aseptically add one K-L filter strip across the
diameter of the plate. Allow the filter strip to sink to the bottom of the
plate or press it down with sterile forceps. Allow the agar to solidify.
Dry the surface of the plates by incubating at 35°C with lid of plate ajar
for 2 hr.
Use: For in vitro toxigenicity testing of Corynebacterium diphtheriae
by the agar diffusion technique. Corynebacterium diphtheriae that pro-
duce toxin form white precipitin lines at approximately 45° angles
from the culture streak line.