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幽门螺旋杆菌Helicobacter pylori ATCC 43504-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

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  • 货  号:幽门螺旋杆菌Helicobacter pylori 43504
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Helicobacter pylori 幽门螺旋杆菌 ATCC 43504

BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心


Designation: NCTC 11637 [JCM 7653, RPH 13487]


Deposited Name: Campylobacter pyloridis Marshall et al.

Product Description: Type strain. Quality control strain for API products. CLSI control strain for

susceptibility testing.


Medium

Medium 18: Trypticase Soy Agar/Broth

Medium 260: Trypticase soy agar/broth with defibrinated sheep blood


Growth Conditions

Temperature: 37°C

Atmosphere: Microaerophilic, 35%

O210%

CO2


Propagation Procedure

1. This organism is shipped frozen in dry ice and should be stored in vapor phase liquid nitrogen until

ready for use. Just prior to use, thaw vial in water bath at approximately 37°C with gentle agitation

until no ice remains. When thawed, a drop of the suspension may be used to do an immediate wet

mount to observe the unique morphology of this organism and verify its viability by checking for

motility.

2. Aseptically transfer the entire thawed suspension into a fresh #18 broth (35

mL). Mix well. This

suspension can now be used to inoculate agar slant(s), plate(s), or the preferred biphasic culture.

Two #260 plates should be inoculated, one for microaerophilic growth and the second for aerobic

growth. No growth should occur on the plate incubated aerobically.

3. To obtain a biphasic culture, add 0.6 mL of the suspension to a #260 slant. The resulting pool at the

bottom of the slant is where the best, most rapid growth will occur.

4. 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 tubes with caps loose.

5. Within 3 days, 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 agar plate. Further subcultures can be made

using the broth pool as the inoculum source. Subcultures to biphasic cultures will require only 24 to

48 hours of incubation for good growth.

Colonies on #260 agar are small, entire, glistening, circular, smooth, and raised.

This is a slowing organism that requires moist conditions for best growth. Growth at the broth/agar interface

of the biphasic slant should occur within 3 days, but only light turbidity will be seen. To observe growth,

examine a wet mount of the broth under phase microscopy. This organism displays motile corkscrew rods

arranged singles.

Growth on agar takes longer than in the biphasic culture. It is essential to use fresh, moist plates. The cells do

not Gram stain well using traditional procedures. For best results, use a basic fuchsin counterstain in place of

the safranin.

Once good growth in 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 "ultralow temperature"

freezer is recommended.

Purified genomic DNA of this strain is available


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