Differentiation of Protein L-Containing and Albumin-BindingPeptostreptococcus magnusIsolates by DNA Amplification, Ribotyping, and Pulsed Field Gel Electrophoresis
Ng1, James
(Department of1Biology and2Microbiology and Immunology, University of Ottawa, Canada)
,
Ng2, Lai-King
(Department of1Biology and2Microbiology and Immunology, University of Ottawa, Canada)
,
Holst3, Elisabet
(3Department of Clinical Microbiology, Lund University Hospital, Lund, Sweden)
,
Dillon1,2, Jo-Anne R.
(Department of1Biology and2Microbiology and Immunology, University of Ottawa, Canada)
AbstractProtein L (L) and albumin-binding (A) proteins may be virulence factors ofPeptostreptococcus magnusisolates. When 32P. magnusisolates from different countries were screened for protein L gene sequences by the polymerase chain reaction (PCR), the only positive isolates were three which had be...
AbstractProtein L (L) and albumin-binding (A) proteins may be virulence factors ofPeptostreptococcus magnusisolates. When 32P. magnusisolates from different countries were screened for protein L gene sequences by the polymerase chain reaction (PCR), the only positive isolates were three which had been previously identified. Protein L-containing (L+A−) or albumin-binding isolates (L−A+) and unrelated isolates (L−A−) were analysed using three genotyping methods not previously used to distinguish strains with these putative virulence factors — PCR-restriction endonuclease analysis (REA) of 16 SrRNA, ribotyping, and macrorestriction endonuclease analysis of DNA by pulsed-field gel electrophoresis (PFGE). PFGE produced the greatest discrimination (ten banding patterns), followed by ribotyping (nine banding patterns) and PCR-REA of 16S rRNA amplicons (two banding patterns). Based on patterns from PFGE and ribotyping, cluster analysis showed that the three L+A−isolates were more closely related to each other than with other isolates. Six of eight L−A+isolates formed two indistinguishable groups of three isolates each, but these two groups were genetically distant from the other two unrelated L−A+isolates. L−A−isolates were not related, either among themselves, or to any other isolates. Thus, L+and A+phenotypes and genotyping methods (PFGE and ribotyping) are useful for differentiatingP. magnusisolates and identifying specific strain types.
AbstractProtein L (L) and albumin-binding (A) proteins may be virulence factors ofPeptostreptococcus magnusisolates. When 32P. magnusisolates from different countries were screened for protein L gene sequences by the polymerase chain reaction (PCR), the only positive isolates were three which had been previously identified. Protein L-containing (L+A−) or albumin-binding isolates (L−A+) and unrelated isolates (L−A−) were analysed using three genotyping methods not previously used to distinguish strains with these putative virulence factors — PCR-restriction endonuclease analysis (REA) of 16 SrRNA, ribotyping, and macrorestriction endonuclease analysis of DNA by pulsed-field gel electrophoresis (PFGE). PFGE produced the greatest discrimination (ten banding patterns), followed by ribotyping (nine banding patterns) and PCR-REA of 16S rRNA amplicons (two banding patterns). Based on patterns from PFGE and ribotyping, cluster analysis showed that the three L+A−isolates were more closely related to each other than with other isolates. Six of eight L−A+isolates formed two indistinguishable groups of three isolates each, but these two groups were genetically distant from the other two unrelated L−A+isolates. L−A−isolates were not related, either among themselves, or to any other isolates. Thus, L+and A+phenotypes and genotyping methods (PFGE and ribotyping) are useful for differentiatingP. magnusisolates and identifying specific strain types.
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