We show here that a combination of both CRISPR-MVLST and PFGE is required to achieve an appropriate discriminatory power for S. Heidelberg. For S. Typhimurium, both subtyping methods independently provide a discriminatory power >0.94. Importantly, as one of the first applications of CRISPR-MVLST to analyze isolates that were part of an outbreak, we were able to cluster two different S. Typhimurium outbreak Selleckchem Ricolinostat strains. Results

Results of CRISPR-MVLST To more accurately determine the discriminatory power of CRISPR-MVLST and PFGE for S. Heidelberg and S. Typhimurium, we subtyped 89 and 86 isolates, respectively, that were obtained from the Pennsylvania Department of Health (Table 1). Among the 175 total isolates analyzed, we identified 29 CRISPR1 alleles, 31 CRISPR2 alleles, 6 fimH alleles and 7 sseL alleles (Table 2). Of these, we found 27, 30, 2 and 4 alleles, respectively, that were novel and not seen in our previous data sets [33]. In total, these alleles defined 58 novel sequence types among the two serovars (Tables 3 and 4). The overwhelming sequence-type diversity among both of these prevalent serovars is provided by genetic variability in the CRISPR

loci, rather than in either fimH or sseL (Figure 2). We found that 88/89 S. Heidelberg isolates had fimH allele 7 and in S. Typhimurium there were two predominant fimH alleles, allele 6 (52/86 isolates) and allele 8 (28/86 isolates). Similarly, in S. Heidelberg, 88/89 isolates bore sseL allele 19 and in S. Typhimurium, 73/86 isolates had sseL allele 15. The polymorphisms between different sseL Smoothened Agonist or fimH alleles arise from the presence of SNPs with the exception of allele 63 that has a single base insertion. No alleles for any of the four markers

were shared among the two different serovars, consistent with previously published studies [32–34]. Table 1 List of 175 S. Heidelberg and S. Typhimurium isolates from the Pennsylvania Department of Health that were analyzed in this study Isolate Sequence type PFGE pattern PA region Isolation date S. Heidelberg         06E00444 HST 7 JF6X01.0022 SE Mar-06 06E00726 SPTLC1 HST 7 JF6X01.0022 SE Jun-06 06E01437 HST 7 JF6X01.0022 SE Aug-06 07E00466 HST 7 JF6X01.0022 SE Apr-07 07E00768 HST 7 JF6X01.0022 NC May-07 07E01405 HST 7 JF6X01.0022 SE Aug-07 07E01505 HST 7 JF6X01.0022 SE Aug-07 08E00753 HST 7 JF6X01.0022 NE Jun-08 08E01373 HST 7 JF6X01.0022 SE Aug-08 09E00637 HST 7 JF6X01.0022 SE Mar-09 09E00701 HST 7 JF6X01.0022 SE Mar-09 09E00750 HST 7 JF6X01.0022 SE Apr-09 09E00782 HST 7 JF6X01.0022 SE Apr-09 09E01149 HST 7 JF6X01.0022 SE May-09 09E01511 HST 7 JF6X01.0022 SE Jun-09 M09019838001A HST 7 JF6X01.0022 SE Aug-09 M10003150001A HST 7 JF6X01.0022 SE Jan-10 M10014816001A HST 7 JF6X01.0022 SE Jun-10 M10016406001A HST 7 JF6X01.0022 SE Jul-10 M10022189001A HST 7 JF6X01.0022 SE Sep-10 M11012103001A HST 7 JF6X01.0022 SW Apr-11 M11017212001A HST 7 JF6X01.0022 SE Jul-11 M11021620001A HST 7 JF6X01.