1887

Abstract

The reported incidence and mortality of -associated disease has increased significantly, which in part is likely to be due to the emergence of a new, highly virulent strain in North America and Europe. This epidemic strain, referred to as BI/NAP1/027, has increased virulence, attributed to overexpression of the two toxin-encoding genes, and , which may be due to truncation of the negative regulator () by a 1 bp deletion. In a previous study of whole-genome comparisons using microarray analysis of 75 isolates, it was noted that the 20 027 strains, which formed a hypervirulent clade, possessed a unique hybridization pattern for the 7 toxin B microarray reporters. This unique pattern was conserved in all of these 027 strains. The pattern was different for the 55 non-027 strains tested. These data, along with the knowledge that 027 strains are toxinotype III (i.e. possess a complete gene of comparable size to toxin reference strain VPI 10463), suggest that the sequence of the N-terminal binding domain of toxin B must be divergent from strain 630 (and the other 55 strains tested). Additionally, these 027 strains had comparable hybridization patterns across the whole microarray, as well as for . Therefore, it was suggested that they share a similar, novel N-terminal binding domain. The aim of this study was to ascertain the sequence variation in from eight characterized BI/NAP1/027 strains. The study confirmed significant sequence variation of from the sequenced strain 630 and slight variation in among the eight 027 strains. These results suggest that toxin B from 027 strains may have a different binding capacity compared with its less-virulent counterparts and may, in addition to the mutated regulator, be responsible for the increased virulence of 027 strains.

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2008-06-01
2020-01-23
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Primers used for amplification and sequencing of regions e–g. [ PDF file] (43 KB)

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Multiple nucleotide sequence alignment of the gene for region e (a), region f (b) and region g (c). The following strains were compared for each region: CD630, BI-1, BI-2, BI-3, BI-9, BI-10, BI-11, BI-13 and BI-14. The primer sequences for amplification of the respective regions are underlined. [ PDF file] (35 KB)

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Multiple sequence alignment of TcdB region e (a), region f (b) and region g (c). BI-1 to BI-3 represent historic non-epidemic fluoroquinolone-sensitive strains, BI-9 to BI-14 represent fluoroquinolone-resistant epidemic strains, BI-9 clustered in the HA1 clade and BI-14 was a hypervirulent clade outlier. [ PDF file] (35 KB)

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