Helicobacter pylori (Marshall et al.) Goodwin et al.
51111™
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Product category
Bacteria
Strain designation
U2-1
Type strain
No
Genome sequenced strain
Yes
Applications
Enteric disease research
Infectious disease research
Product format
Freeze-dried
General
Specific applications
Enteric Research
Preceptrol
No
Characteristics
Comments
Urease-negative variant of ATCC 49503
Handling information
Medium
ATCC Medium 260: Trypticase soy agar/broth with defibrinated sheep blood
Temperature
37°C
Atmosphere
Microaerophilic
Handling procedure
1. Open vial according to enclosed instructions. Rehydrate
contents of vial with 0.5 ml of Trypticase Soy Broth.
2. To obtain a biphasic culture, add 0.4 ml of the suspension
to a #260 slant. Add remaining 0.1 ml of the suspension to a #260 plate and streak for isolation.
3. Incubate at 37°C under microaerophilic conditions using
an anaerobe jar with an active catalyst and a microaerophilic gas generator pack, or other acceptable method, to obtain microaerophilic conditions. Incubate slant with cap loose.
4. Within three days of incubation, good growth should be obtained in the broth pool at the bottom of the slant. Additional incubation may be required for colonies to appear on the plate. Further subcultures can be made using broth pool as the inoculum source.
4.
Handling notes
This is a slow growing organism that requires moist conditions for best growth. Growth at the broth/agar interface of the biphasic slant should occur within three days, but little turbidity will be seen. To observe growth, examine a wet mount of the broth under phase microscopy. The organism is a medium size, regular to slightly curved, motile bacillus. Motility is usually observed only in young cultures. The presence of spheroid cells indicates that viability is being lost either due to age or too much exposure to oxygen.
Growth on agar takes longer than with the biphasic culture. Colonies are small, circular, entire, convex, smooth, and gray. Once good growth is present, these organisms tend to lose viability, especially if exposed to air for lengthy periods. Viability also decreases with repeated subculturing. The cells do not Gram stain well using traditional procedures. To obtain the best results, use a basic fuchsin counterstain in place of the safranin.
Once good growth is obtained, transfer or freeze the culture. Adding an equal amount of 20% sterile glycerol to pooled broth from several biphasic slants, followed by freezing in liquid nitrogen or "ultra-low temperature" freezer is recommended.
Additional information on this culture is available on the ATCC web site at www.atcc.org.
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