title: Development of Prevotella nigrescens-specific PCR primers based on the nucleotide sequence of a Pn23 DNA probe.
abstract: A previous study reported the cloning of a putative Prevotella nigrescens-specific DNA probe, Pn23, using random shotgun method. The present study evaluated the species-specificity of Pn23 for P. nigrescens using the clinical strains of Prevotella intermedia and P. nigrescens to develop P. nigrescens-specific polymerase chain reaction (PCR) primers. Southern blot analysis showed that the DNA probe, Pn23, detected only the genomic DNA of P. nigrescens strains. PCR showed that the two sets of PCR primers, Pn23-F1/Pn23-R1 and Pn23-F2/Pn23-R2, had species-specificity for P. nigrescens. Interestingly, the two sets of PCR primers, Pn23-F6/Pn23-R6 and Pn23-F7/Pn23-R7, had strain-specificity for P. nigrescens ATCC 33563. The detection limits of the four primer sets were 40 or 4 pg of the purified genomic DNA of P. nigrescens ATCC 33563. These results suggest that the DNA probe, Pn23, and the two sets of PCR primers, Pn23-F1/Pn23-R1 and Pn23-F2/Pn23-R2, can be useful for the detection of P. nigrescens in the molecular epidemiological studies of oral infectious diseases.
authors.affiliation: Department of Oral Biochemistry, School of Dentistry, Chosun University, 375 Seosuk-Dong, Dong-Gu, Gwangju 501-759, Republic of Korea.
authors.forename: Min Jung
authors.initials: MJ
authors.lastname: Kim
authors.forename: Young-Seok
authors.initials: YS
authors.lastname: Lee
authors.forename: Jae-Yoon
authors.initials: JY
authors.lastname: Park
authors.forename: Joong-Ki
authors.initials: JK
authors.lastname: Kook
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D001483:Base Sequence; D015139:Blotting, Southern; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D016905:Gram-Negative Bacterial Infections / Q000175:diagnosis; D006801:Humans; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D018713:Prevotella intermedia / Q000235:genetics; D045242:Prevotella nigrescens / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D015345:Oligonucleotide Probes