Clostridioides difficile (Prevot) Lawson et al.
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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
Anaerobic conditions for transfer may be obtained by either of the following:
Anaerobic conditions for incubation may be obtained by any of the following:
Anaerobe Systems Brucella Blood Agar Plates (AS-111) can be used to analyze colony morphology and purity.
No growth should occur on the aerobic blood plate. Turbid growth is evident is broth at 24 hours. Subsequent transfers may reach peak viability in 6 to 12 hours.
The presence of tcdA and tcdB genes is confirmed by PCR. The binary toxin gene cdt is not amplified by PCR.
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Lemee L, et al. Multiplex PCR targeting tpi (triose phosphate isomerase), tcdA (Toxin A), and tcdB (Toxin B) genes for toxigenic culture of Clostridium difficile. J. Clin. Microbiol. 42(12): 5710-5714, 2004. PubMed: 15583303
Rupnik, M et al. A novel toxinotyping scheme and correlation of toxinotypes with serogroups of Clostridium difficile isolates. J. Clin. Microbiol. 36(8): 2240-2247, 1998. PubMed: 9665999
Fawley WN, Wilcox MH; on behalf of the Clostridium difficile Ribotyping Network for England and N. Ireland (CDRN). An enhanced DNA fingerprinting service to investigate potential Clostridium difficile infection case clusters sharing the same PCR-ribotype. J. Clin. Microbiol. 49(12): 4333-4337, 2011. PubMed: 21956986