Differentiation of Protein L-Containing and Albumin-Binding Peptostreptococcus magnus Isolates by DNA Amplification, Ribotyping, and Pulsed Field Gel Electrophoresis

Author: Ng J.   Ng L.K.   Holst E.   Dillon J.A.R.  

Publisher: Academic Press

ISSN: 1075-9964

Source: Anaerobe, Vol.2, Iss.2, 1996-02, pp. : 95-102

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Abstract

Protein L (L) and albumin-binding (A) proteins may be virulence factors of Peptostreptococcus magnus isolates. When 32 P. magnus isolates 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 differentiating P. magnus isolates and identifying specific strain types.

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