The results of the tests for examining intragenic recombination (

The results of the tests for examining intragenic recombination (recombination within the sequence of a gene) are summarised in Table  2. For each test the number of loci that were positive for recombination is recorded. For RDP at least two of the individual tests in the suite had to Selleck RAD001 be positive in order for the locus to be

scored positive overall. Table 2 Number of loci positive for recombination by the Sawyer’s run test and RDP suite   Sawyer’s run test RDP tests Staphylococcus aureus (Clonal) 0 loci 1 locus Streptococcus pneumoniae (Intermediate) 3 loci 4 loci Neisseria menigitidis (Panmictic) 7 loci 6 loci Legionella pneumophila 1 locus 2 loci Both the Sawyer’s run test and RDP show L. pneumophila has an intermediate rate of

intragenic recombination when compared with other bacterial species. Overall the collected evidence from this and several previous studies [12–14, 16, 17, 23] strongly suggest that L. pneumophila is not a purely clonal organism but also undergoes significant recombination. The results presented here suggest that L. pneumophila retains evidence for a clonal vertical inheritance of genetic material whilst also demonstrating strong evidence of recombination by horizontal transfer of genetic loci. Although there was some evidence for recombination within the SBT genes, the frequency was low and this indicates selleck chemical that new alleles are most likely to be generated by point mutations 2-hydroxyphytanoyl-CoA lyase rather than recombination. The signal from vertical inheritance of genetic material through clonal lineages is still evident when examining the genetic information contained from seven L. pneumophila loci. However it is also clear that recombination happens often enough so that it is a significant force in shaping the population structure. This does not alter the utility of SBT as a means to discriminate between isolates of L. pneumophila, particularly for outbreak investigation, since the results indicate that it is far from being a panmictic organism. Although we

cannot infer a rate of recombination from this study, the relatively low frequency of recombination suggests that recombination would be unlikely to take place in the timescale of an outbreak and therefore the ST of isolates involved in an outbreak is also unlikely to change. Sequence Based Typing analysis: Clustering Since the ultimate aim of this work was to find a practical way to cluster L. pneumophila isolates, a method of determining which clustering method resulted in the most accurate sub-groups was required. Given that the recombination analysis above indicates that clonal vertical inheritance plays a major role in the evolution of L. pneumophila, a phylogenetic tree based on the genetic distance between the concatenated sequences from the SBT loci will provide an approximate representation of the evolutionary history.

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