title: Development of Prevotella intermedia-specific PCR primers based on the nucleotide sequences of a DNA probe Pig27.
abstract: The aim of this study was to develop Prevotella intermedia-specific PCR primers based on the P. intermedia-specific DNA probe. The P. intermedia-specific DNA probe was screened by inverted dot blot hybridization and confirmed by Southern blot hybridization. The nucleotide sequences of the species-specific DNA probes were determined using a chain termination method. Southern blot analysis showed that the DNA probe, Pig27, detected only the genomic DNA of P. intermedia strains. PCR showed that the PCR primers, Pin-F1/Pin-R1, had species-specificity for P. intermedia. The detection limits of the PCR primer sets were 0.4pg of the purified genomic DNA of P. intermedia ATCC 49046. These results suggest that the PCR primers, Pin-F1/Pin-R1, could be useful in the detection of P. intermedia as well as in the development of a PCR kit in epidemiological studies related to periodontal diseases.
authors.affiliation: Department of Oral Biochemistry, School of Dentistry, Chosun University, 375 Seosuk-Dong, Dong-Ku, Gwangju 501-759, Republic of Korea.
authors.forename: Min Jung
authors.initials: MJ
authors.lastname: Kim
authors.forename: Kyung Hwan
authors.initials: KH
authors.lastname: Hwang
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*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018713:Prevotella intermedia / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D004269:DNA, Bacterial; D015345:Oligonucleotide Probes