RAPD fingerprinting for the distinction of Prevotella intermedia sensu stricto from Prevotella nigrescens
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Published on: October 23, 2011
1Groupe de Recherche en Ecologie Buccale, Faculté de Médecine Dentaire, Université Laval, Québec, Canada, G1K 7P4.
Abstract:
A collection of 70 oral strains including reference strains and clinical isolates identified as Prevotella intermedia sensu lato was constituted to cover a large clinical and geographical diversity. Electrophoresis of the enzyme malate dehydrogenase allowed the identification of the 70 study strains as Prevotella intermedia sensu stricto (n= 36), Prevotella nigrescens (n= 31) and three unclassified strains. By using four primers, DNA fingerprints were generated from 20 strains as random amplified polymorphic DNA (RAPD). Matching co-migrating amplicon positions by pairwise comparison allowed the clustering of the fingerprints as two groups coincident with the P. intermedia/P. nigrescens assignment by enzyme electrophoresis of malate dehydrogenase. Our data suggest that isolates identified asP. intermedia sensu lato by conventional criteria can be speciated asP. intermedia sensu stricto or P. nigrescens by RAPD fingerprinting.
Insights
Random amplified polymorphic DNA (RAPD) fingerprinting effectively differentiates Prevotella intermedia sensu stricto and Prevotella nigrescens. This molecular method aids in accurate speciation of oral Prevotella strains previously identified broadly.
Area of Science:
- Microbiology
- Molecular Biology
- Bacteriology
Background:
- Prevotella intermedia sensu lato is a complex group of oral bacteria.
- Accurate identification and speciation are crucial for understanding their clinical and epidemiological roles.
Purpose of the Study:
- To evaluate the utility of Random Amplified Polymorphic DNA (RAPD) fingerprinting for differentiating Prevotella intermedia sensu stricto and Prevotella nigrescens.
- To compare RAPD with enzyme electrophoresis for bacterial speciation.
Main Methods:
- Seventy oral Prevotella strains were analyzed using malate dehydrogenase enzyme electrophoresis.
- DNA fingerprinting was performed on 20 selected strains using Random Amplified Polymorphic DNA (RAPD) with four primers.
- RAPD fingerprints were compared to cluster strains based on DNA amplicons.
Main Results:
- Enzyme electrophoresis identified 36 strains as P. intermedia sensu stricto, 31 as P. nigrescens, and 3 unclassified.
- RAPD fingerprinting generated distinct patterns that clustered into two groups.
- These RAPD clusters corresponded precisely with the speciation obtained by enzyme electrophoresis.
Conclusions:
- Random Amplified Polymorphic DNA (RAPD) fingerprinting is a reliable and effective method for differentiating Prevotella intermedia sensu stricto and Prevotella nigrescens.
- RAPD offers a valuable molecular tool for the accurate speciation of oral Prevotella isolates previously identified under the broader P. intermedia sensu lato classification.
