Int J Oral Biol.  2010 Mar;35(1):13-19.

Detection of Prevotella intermedia and Prevotella nigrescens using Pn17 and Pn34 DNA Probes

• Park CH
• Kim PS
• Kim HS
• Min JB
• Hwang HK
• Jang HS
• Cho KW
• Baek DH
• Kook JK

Abstract

The DNA probes Pn17 and Pn34 were evaluated for their ability to specifically detect clinical strains of P. intermedia and P. nigrescens from a Korean population by dot blot hybridization. These probes were sequenced by extension termination and their specificity was determined by Southern blot analysis. The results revealed that the Pn17 sequence (2,517 bp) partially encodes an RNA polymerase beta subunit (rpoB) and that Pn34 (1,918 bp) partially encodes both rpoB (1-169 nts) and the RNA polymerase beta subunit (rpoB'; 695-1918 nts). These probes hybridized with both HindIII- and PstI-digested genomic DNAs from the strains of P. intermedia and P. nigrescens used in this study. Interestingly, each of the hybrid bands generated from the HindIII-digested genomic DNAs of the two bacterial species could be used to distinguish between them via restriction fragment length polymorphism. These results thus indicate that Pn17 and Pn34 can simultaneously detect P. intermedia and P. nigrescens.

Keyword

DNA probes Pn17 and Pn34; Prevotella intermedia; Prevotella nigrescens

MeSH Terms

Blotting, Southern
Chimera
DNA
DNA Probes
DNA-Directed RNA Polymerases
Polymorphism, Restriction Fragment Length
Prevotella
Prevotella intermedia
Prevotella nigrescens
Sensitivity and Specificity
DNA
DNA Probes
DNA-Directed RNA Polymerases