03 99 cd38 7 811821-34 2 2 0.03 100 a Accesstion number in Genbank is AM180355. b Identified previously by Marsh et al. [13] and van den Berg et al. [14]. c This locus contains incomplete repeat and is denoted by the size of array. Capillary gel electrophoresis-based PCR ribotyping Of the 142 isolates, capillary gel electrophoresis-based PCR-ribotyping identified 57 independent types, including 32 singletons. The most common types were R45, R4, R10, R14, and R17 (UK017), containing 7, 17, 11, 11 and 9 isolates, respectively (Figure 1). The R27 (UK 027) virulent type was not found among the local strains. learn more Figure 1 Comparison of PCR riboytpe and MLVA groups for 142 C. difficile isolates. Dendrogram

is based on UPGMA analysis of capillary electrophoresis-based PCR ribotyping, and the vertical line is the cutoff point for identifying PCR-ribotype groups. Corresponding PCR-ribotype Volasertib order groups, MLVA34 groups, MLVA10 groups, and number of isolates are shown. MLVA groups are identified by minimum-spanning tree: one group is defined by MLVA type with less than two loci difference.

Dendrogram based on PCR ribotyping A phylogenetic dendrogram based on the PCR-ribotypes was constructed using the 142 C. difficile isolates (Figure 1). Of the 142 isolates, PCR-ribotype, MLVA34, and MLVA10, identified 57 types, 47 groups, and 45 groups, respectively. The PCR-ribotype was more discriminatory than the two MLVA groups (Figure 1). Using a Dichloromethane dehalogenase threshold of >83% similarity for defining PCR-ribotype groups, all isolates were able to be divided into 47 PCR-ribotype groups, including 22 singletons. Over 87% (41/47) of the PCR-ribotype groups

were specifically recognized in the MLVA34 and MLVA10 groups. However, PCR-ribotype groups 39 and 25 were recognized together as one by both MLVA groups, with the fingerprints for these isolates sharing a 70% similarity (a four-band difference). In addition, PCR ribotype groups 26 and 49 were also identified as one by the two MLVA groups, with the fingerprints of these two isolates sharing a 78% similarity. Furthermore, PCR ribotype groups 8 and 23 were also seen as one by the two MLVA groups, with the fingerprint of these isolates sharing an 82% similarity. Taken together, these results shows that this discordance, the lack of one to one identification between PCR ribotypes and MLVA groups, mainly occurred when PCR-ribotypes shared >83% similarity. Congruence between groups of the PCR ribotype and MLVA MLVA panels with slightly limit allelic diversity PLX3397 mouse generated groups highly congruent with PCR ribotyping (Table 2). To determine the most congruent groupings between MLVA panels and PCR-ribotype groups, groupings of MLVA panels consisting of VNTR loci with high to low allelic diversity were compared with the PCR-ribotype groups. MLVA34, MLVA12, and MLVA10 generated partitions (47, 45, and 45, respectively) and allelic diversity (0.959, 0.957, and 0.957, respectively) similar to those identified by PCR ribotyping (Table 2).