LAB 140

Description

A selective medium for the isolation of Helicobacter pylori, causative agent of chronic gastritis.

This campylobacter-like organism was described in 1983 colonising the gastric mucosa, a site previously thought to be sterile due to the low PH. The high level of Urease production by this organism appears to be the major pathogenicity factor enabling it to withstand the strongly acidic environment.

Helicobacter Pylori Medium is a modification of CCDA Medium for the isolation of Campylobacter spp. Formulated by Bolton et al at Preston Public Health Laboratory, it incorporates a rich agar base supplemented with horse serum to promote optimum growth, and Vancomycin, Cefsulodin, and Amphotericin as selective agents.

Method for reconstitution

Weigh 45.0 grams of powder and disperse in 1 litre of deioni water. Soak for 10 minutes, swirl to mix and sterilise at 121˚C fo minutes. Cool to 47˚C and add 100ml of Horse Serum, and 2 vial VCA supplement X040. Mix well and pour, continuing to mix wh pouring to keep the charcoal in suspension.

Appearance: Black Agar

pH: 7.4 ± 0.2

NCIMB 5008

Storage of prepared medium: Plates - up to 7 days at 2-the dark.

Inoculation: Surface streaking for single colonies.

Incubation: 37 ˚C for 72 hours.

References

Bolton et al. Public Health Laboratory, Preston. Personal communication.