Application Guide - QIAxcel Advanced

88 QIAxcel Advanced Application Guide 10/2016 A B References 1. Braun, V., Hundsberger, T., Leukel, P., Sauerborn, M., and von Eichel-Streiber, C. (1996) Definition of the single integration site of the pathogenicity locus in Clostridium difficile. Gene 182, 29. 2. Von Eichel-Streiber, C. Laufenberg-Feldmann, R., Sartingen, S., Schulze, J., and Sauerborn, M. (1992) Comparative sequence analysis of the Clostridium difficile toxins A and B. Mol. Gen. Genet. 233, 260. 3. O‘Neill, G.L., Ogunsola, F.T., Brazier, J.S., and Duerden, B.I. (1996) Modification of a ribotyping method for application as a routine typing scheme for Clostridium difficile. Anaerobe 2, 205. 4. Stubbs, S.L.J., Brazier, J.S., O’Neill, G.L., and Duerden, B.I. (1999) PCR targeted to the 12S-23S rRNA gene intergenic spacer region of Clostridium difficile and construction of a library consisting of 116 different PCR ribotypes. J. Clin. Microbiol. 37, 461. 5. Gonçalves, C., Decré, D., Barbut, F., Burghoffer, B., and Petit, J.C. (2004) Prevalence and characterization of a binary toxin from Clostridium difficile. J. Clin. Microbiol. 42, 1933. 6. Spigaglia, P. and Mastrantonio, P. (2002) Molecular analysis of the pathogenicity locus and polymorphism in the putative negative regulator of toxin production (TcdC) among Clostridium difficile clinical isolates. J. Clin. Microbiol. 40, 3470. 7. Indra, A. et al. (2008) Characterization of Clostridium difficile isolates using capillary gel electrophoresis-based PCR ribotyping. J. Med. Microbiol. 57, 1377. 8. van den Berg, R.J., Schaap, I., Templeton, K.E., Klaassen, C.H.W., and Kuijper, E.J. (2007) Typing and subtyping of Clostridium difficile isolates by using multiple-locus variable-number tandem-repeat analysis. J. Clin. Microbiol. 45, 102. 9. Fawley, W.N. et al. (2008) Use of highly discriminatory fingerprinting to analyze clusters of Clostridium difficile infection cases due to epidemic ribotype 027 strains. J. Clin. Microbiol. 46, 954. Figure 2. C. difficile genotyping. A. Partial genotyping of a C. difficile 027 ribotype sample. B01: tcdC control; B02: tcdC with deletion; B03: cdtA/B; B04: tcdA/B. B. Overlay of electropherogram views of B01 (206.8 bp, blue) and B02 (188.4 bp, green) lanes from part C. demonstrating the 18 bp tcdC deletion. Peaks for the upper and lower alignment marker are indicated in red. Conclusions The QIAxcel system proved to be suitable for C. difficile ribotyping applications and toxin-gene detection. Due to significantly shorter run times in comparison to conventional methods, the system has the potential to reduce the cost of PCR ribotyping by drastically reducing the hands-on time. Standardized automated processing facilitates inter- laboratory data exchange without the need for cumbersome standardization of equipment, reagents, and operating procedures.

Pages Overview