In the latest years an increasing number of genomes have been sequenced paving the path for genomics-based approaches. For P. gingivalis genome sequences of the virulent strain W83 and the less-virulent strain ATCC33277 have become available [28, 29]. Comparative genomic hybridization (CGH) analysis using microarrays of these well-described bacterial strains could yield new insights in the virulence mechanisms of P. gingivalis. A recent study reported on the CGH analysis of several P. gingivalis strains to describe the genetic buy PF-6463922 variety among them [30]. In this study we analyzed the genetic contents of representative strains of each of the seven capsular serotypes (Table 1): W83 (K1), HG184
(K2), ATCC53977 (K3), ATCC49417 (K4), HG1690 (K5), HG1691 (K6), 34-4 (K7). We also included the non-encapsulated strain FDC381 (K-) in the CGH analysis to compare with each of the encapsulated strains. Strain FDC381 does however express a non-CPS anionic extracellular polysaccharide as do the other strains [31]. The strains were classified into three virulence levels as determined by using a subcutaneous mouse infection model [18, 32]. Although not an optimal measure for the ability to cause periodontitis, this classification has long been used [33] and proven useful in studying virulence determinants [34–37]. Table 1 P. gingivalis strains used in this study Strain Capsular serotype Origin Virulencec W83a K1 Clinical
specimen High HG184 K2 Periodontitis
patient Medium HG1025 K3 Periodontitis patient with diabetes Wortmannin mellitus High ATCC49417 K4 Advanced adult periodontitis patient High HG1690 K5 37-year-old male periodontitis patient High HG1691 K6 28-year-old female periodontitis patient Medium 34-4 K7 Severe periodontitis patient Low FDC381b else K- Adult periodontitis patient Low a A kind gift of H. N. Shah (NCTC, London, UK) b A kind gift of S. S. Socransky (The Forsyth selleck Institute, Boston, MA, USA) c As determined in a subcutaneous mouse infection model [18, 32] Triplicate hybridization experiments and three types of analysis, 1) aberrant gene calling, 2) breakpoint analysis and 3) absent gene calling, have been performed for optimal use of the new genetic information. The careful design of the experiment and the thorough analysis of the data lead to a high resolution data set, yielding more detailed information on the genetic differences between strains than has been shown before. In this study we initiate the description of a core-gene set of P. gingivalis allowing a more focused search for potential important virulence factors. Results and discussion Microarray performance and data interpretation The P. gingivalis version 1 microarray from the PFGRC used in this study has been used in several studies before [30, 38] and consists of 1907 probes and 500 negative control probes (Arabidopsis thaliana) printed in four replicates.
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