Polymerase chain reaction analysis of the clonality of Porphyromonas gingivalis and collagenase gene.
OBJECTIVE
To investigate the genotypic characterization of Porphyromonas gingivalis (Pg) and the heterogeneity of a potential virulence factor-PrtC.


METHODS
Arbitrarily primed polymerase chain reaction (AP-PCR) was applied to 80 Pg strains isolated from 24 unrelated Chinese periodontitis patients. PCR reaction was used to detect a fragment of the collagenase gene (PrtC gene). To evaluate the sequence heterogeneity of the Pg PrtC genes, sequence analysis of four PrtC gene of clinical isolates was performed.


RESULTS
Random primer OPA-05 and OPA-17 distinguished 7 AP-PCR profiles (I through VII). The majority of the strains belonged to type VII which accounted for 25.8%. A 548bp fragment of PrtC gene was detected from 24 clinical strains. The PCR products were verified by the restriction endonucleases PstI and PvuI. Sequence analysis showed 4 PrtC genes were heterogeneous in their nucleotide composition and differed from reference strain Pg 53977.


CONCLUSIONS
The results demonstrated genetic diversity existed among these clinical strains isolated from Chinese periodontitis patients and the PrtC genes are heterogeneous in their nucleotide sequence.
Jia
Xiaozhen
X
Department of Periodontics, School of Stomatology, Wuhan University, Wuhan 430079, China.
Fan
Mingwen
M
Bian
Zhuan
Z
Chen
Zhi
Z
Li
Chengzhang
C
D000328:Adult; D017364:Collagenases / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005260:Female; D018740:Genetic Heterogeneity; D006801:Humans; D008297:Male; D008875:Middle Aged; D010518:Periodontitis / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods; D016966:Porphyromonas gingivalis / Q000235:genetics* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA
D004269:DNA, Bacterial; D017364:Collagenases