Identification of oral Actinomyces species using DNA probes.
Identification of oral Actinomyces species using DNA probes.
复制标题
使用 DNA 探针鉴定口腔放线菌种类。
DOI：
10.1034/j.1399-302x.1999.140410.x
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发表时间：
1999
影响因子：
--
通讯作者：
Socransky,SS
中科院分区：
文献类型：
--
作者：
Ximénez-Fyvie,LA;Haffajee,AD;Martin,L;Tanner,A;Macuch,P;Socransky,SS
OralActinomycescomprise a major segment of both the supra‐ and subgingival microbiota; however, little is known about the distribution of individual species in different sites or clinical conditions. The purpose of the present investigation was to develop DNA probes for suggested species and genotypes of oralActinomyces. Whole genomic DNA probes to 12 human oral species and/or serotypes were labeled with digoxigenin and used to seek cross‐reactions among the taxa using the checkerboard DNA‐DNA hybridization assay. TheActinomycesformed three distinct groups: 1)Actinomyces georgiae,Actinomyces meyeriandActinomyces odontolyticusserotypes I and II; 2)Actinomyces viscosusandActinomyces naeslundiiserotypes I, II, III and WVA 963; and 3)Actinomyces gerencseriaeandActinomyces israelii. Cross‐reactions among taxa were detected and minimized by increasing the temperature of the post‐hybridization high‐stringency wash to 80°C. Despite the elevation in high stringency wash temperature, cross‐reactions among strains of theA. naeslundii/A. viscosusgroup persisted. Probes for two of the three currently recognized genospecies in this group were prepared by removing the DNA in common between cross‐reacting species using subtraction hybridization and polymerase chain reaction. Nine species and genospecies could be clearly separated by a combination of whole genomic and subtraction hybridization probes and by increasing the high‐stringency wash temperature. A total of 195 fresh isolates ofActinomyceswere grouped in a blind study using DNA probes and separately by SDS‐PAGE protein profiles. Concordance between the two methods was 97.3%. The probes and hybridization conditions were tested for their ability to detect theActinomycesspecies and genospecies in samples of supragingival and subgingival plaque from periodontitis subjects using checkerboard DNA‐DNA hybridization. The probes detected the species in samples of supragingival and subgingival plaque. We concluded that whole genomic and subtraction hybridization DNA probes facilitate the detection and enumeration of species and genospecies ofActinomycesin plaque samples.
