RESUMEN
Genetic analysis of 31 clinical strains of Porphyromonas gingivalis isolated from nine subjects, 2-6 strains per subject, was performed by Southern hybridization. Chromosomal DNA was extracted by the method of Moncla et al. [1] and digested to completion with restriction endonucleases PstI, ClaI and BglI. The DNA fragments were separated electrophoretically on agarose gels, transferred to nylon membranes and hybridized to the non-radioactively labelled plasmid pKK 3535 which contains the rmB ribosomal RNA operon of the Escherichia coli chromosome. Of the three enzymes, BglI was the most suitable for the genetic analysis of P. gingivalis. With this enzyme, the intra-individual strains were shown to be identical in eight of the nine subjects, whereas inter-individual strains were different.
Asunto(s)
Porphyromonas gingivalis/genética , Infecciones por Bacteroides/microbiología , ADN Bacteriano/análisis , ADN Bacteriano/genética , ADN Ribosómico/genética , Desoxirribonucleasas de Localización Especificada Tipo II , Escherichia coli/genética , Genotipo , Humanos , Plásmidos , Porphyromonas gingivalis/aislamiento & purificación , Especificidad de la EspecieRESUMEN
The transmission of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and mutans streptococci was studied between 4 married couples who suffered from advanced periodontitis. Of the 20 couples investigated, the 4 in which both spouses harbored A. actinomycetemcomitans and P. gingivalis were chosen for the transmission study. Three of these couples also harbored mutans streptococci. A. actinomycetemcomitans isolates (8-24 per subject) and mutans streptococcal isolates (5-23 per subject) were serotyped by immunodiffusion technique. For ribotyping, chromosomal DNA from A. actinomycetemcomitans isolates (4-5 per subject) and mutans streptococcal isolates (4-11 per subject) was digested with restriction endonucleases ClaI or BglI and HindIII or SmaI, respectively. P. gingivalis isolates (2-15 per subject) were ribotyped by using ClaI, BglI and SmaI. The blotted restriction fragments were hybridized to the plasmid pKK3535, which contains the rRNA operon of the E. coli chromosome. The spouses in 2 couples shared the same sero- and ribotypes of A. actinomycetemcomitans and S. mutans. P. gingivalis ribotypes were identical in 2 couples. The result suggests transmission of oral bacteria between spouses.