fimA genotypes and PFGE profile patterns in Porphyromonas gingivalis isolates from subjects with periodontitis.
OBJECTIVE
Porphyromonas gingivalis is frequently identified to type by evaluation of fimA polymorphisms and less often by pulsed-field gel electrophoresis (PFGE) because of the technical intricacies of PFGE. To compare these techniques, we genotyped P. gingivalis clinical isolates as to (i) their fimA type and (ii) their whole genome restriction profile (PFGE analysis).


METHODS
Thirty-two P. gingivalis strains were isolated from 16 unrelated periodontitis patients. Two strains were isolated from each patient. Strains were subjected to a fimA-typing polymerase chain reaction (PCR) assay. Strains that could not be typed by PCR were submitted to sequencing of the entire fimA gene. The PFGE profiles of clinical strains were compared using bioinformatic analysis.


RESULTS
Seven of the 32 isolates were not typeable by PCR and so their entire fimA gene was sequenced. The sequencing identified each strain as belonging to a single fimA type. In one case, sequencing of the fimA gene did not agree with the result obtained using fimA PCR typing. With the exception of one patient, each patient presented isolates bearing the same fimA type. However, in three patients, isolates with the same fimA type presented different PFGE pulsotypes.


CONCLUSIONS
The P. gingivalis typing using fimA PCR has limitations in typeability and discriminatory power. A typing technique for P. gingivalis that is easy to perform but that presents adequate typeability and discriminatory power is needed if we want to better understand the epidemiology of periodontal disease.
Perez-Chaparro
P J
PJ
Equipe de Microbiologie, UPRES-EA 1254, Université de Rennes 1, Université Européenne de Bretagne, Rennes, France.
Rouillon
A
A
Minet
J
J
Lafaurie
G I
GI
Bonnaure-Mallet
M
M
D015373:Bacterial Typing Techniques; D016866:Bacteroidaceae Infections / Q000382:microbiology*; D002999:Clone Cells / Q000145:classification; D016521:Electrophoresis, Gel, Pulsed-Field; D036781:Fimbriae Proteins / Q000145:classification* / Q000235:genetics; D016680:Genome, Bacterial / Q000235:genetics; D005838:Genotype; D006801:Humans; D010518:Periodontitis / Q000382:microbiology*; D010802:Phylogeny; D018933:Pili, Sex / Q000145:classification* / Q000235:genetics; D016966:Porphyromonas gingivalis / Q000145:classification* / Q000302:isolation & purification
C063055:fimbrillin; D036781:Fimbriae Proteins