Mättö J, Saarela M, von Troil-Lindén B, Alaluusua S, Jousimies-Somer H, Asikainen S
Institute of Dentistry, University of Helsinki, Finland.
Oral Microbiol Immunol. 1996 Dec;11(6):395-401. doi: 10.1111/j.1399-302x.1996.tb00201.x.
The distribution and the genetic similarity of Prevotella intermedia and Prevotella nigrescens in saliva and in subgingival samples recovered from the same subject were studied in 16 subjects with different periodontal status. The isolates (4 salivary and 4 subgingival P. intermedia/nigrescens group isolates per subject) were identified to species level by hybridization with species-specific oligonucleotide probes, and the clonal analysis was performed using arbitrarily primed polymerase chain reaction (AP-PCR) (all isolates) and ribotyping (isolates from 5 subjects). In addition, the applicability of AP-PCR in differentiating between P. intermedia and P. nigrescens species was tested using 18 P. intermedia and 20 P. nigrescens isolates from 34 subjects. P. intermedia was detected in 7 and P. nigrescens in 14 of the 16 subjects. In all subjects the same species was found both in saliva and in subgingival plaque. In 15 of the 16 subjects, similar AP-PCR types of P. intermedia and/or P. nigrescens between salivary and subgingival samples were found. The salivary and subgingival isolates that were similar by AP-PCR were indistinguishable also by ribotyping. The AP-PCR analysis revealed a P. intermedia or P. nigrescens species-specific AP-PCR product in most isolates. This study indicates that both P. intermedia and P. nigrescens were found both in salivary and in subgingival samples, and both sampling sites within the same individual were usually colonized with identical AP-PCR types of the species. Thus, in addition to a subgingival sample a salivary sample seems to be suitable for detection and clonal analysis of these species. The AP-PCR method proved to be a simple method applicable for differentiation and clonal analysis of P. intermedia and P. nigrescens.
在16名患有不同牙周状况的受试者中,研究了中间普氏菌和变黑普氏菌在同一受试者唾液和龈下样本中的分布及遗传相似性。通过与物种特异性寡核苷酸探针杂交将分离株(每位受试者采集4株唾液和4株龈下中间普氏菌/变黑普氏菌群分离株)鉴定到种水平,并使用任意引物聚合酶链反应(AP-PCR)(所有分离株)和核糖体分型(来自5名受试者的分离株)进行克隆分析。此外,使用来自34名受试者的18株中间普氏菌和20株变黑普氏菌分离株测试了AP-PCR在区分中间普氏菌和变黑普氏菌物种方面的适用性。16名受试者中,7名检测到中间普氏菌,14名检测到变黑普氏菌。在所有受试者的唾液和龈下菌斑中均发现了相同的物种。16名受试者中有15名在唾液和龈下样本中发现了相似的中间普氏菌和/或变黑普氏菌AP-PCR类型。通过AP-PCR相似的唾液和龈下分离株在核糖体分型中也无法区分。AP-PCR分析在大多数分离株中揭示了中间普氏菌或变黑普氏菌物种特异性的AP-PCR产物。本研究表明,中间普氏菌和变黑普氏菌在唾液和龈下样本中均有发现,并且同一个体内的两个采样部位通常被相同AP-PCR类型的该物种定殖。因此,除了龈下样本外,唾液样本似乎也适用于这些物种的检测和克隆分析。AP-PCR方法被证明是一种适用于中间普氏菌和变黑普氏菌鉴别及克隆分析的简单方法。