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Porphyromonas gingivalis as a Model Organism for Assessing Interaction of Anaerobic Bacteria with Host Cells
Published on: December 17, 2015
fimA genotypes and multilocus sequence types of Porphyromonas gingivalis from patients with periodontitis
Morten Enersen1, Ingar Olsen, Øyvind Kvalheim
1Institute of Oral Biology, Faculty of Dentistry, University of Oslo, P.O. Box 1052, NO-0316 Oslo, Norway. morteene@odont.uio.no
Abstract:
Fimbriae are important virulence factors of pathogenic bacteria, facilitating their attachment to host and bacterial cells. In the periodontal pathogen Porphyromonas gingivalis, the fimA gene is classified into six types (genotypes I, Ib, II, III, IV, and V) on the basis of different nucleotide sequences, with fimA genotypes II and IV being prevalent in isolates from patients with periodontitis. The aims of this study were to examine the distribution of fimA genotypes in a collection of 82 P. gingivalis isolates from adult periodontitis patients of worldwide origin and to investigate the relationship between the fimA genotypes and the sequence types (STs), as determined by multilocus sequence typing (MLST), of the isolates. The fimA gene was amplified by PCR with primer sets specific for each genotype. The STs of all strains were assigned according to the MLST database for P. gingivalis (www.pubmlst.org/pgingivalis). The 82 strains showed extensive genetic diversity and were assigned to 69 STs. Only isolates with closely related STs harbored the same fimA genotype. Twenty-eight (34.1%) strains harbored fimA genotype II, while only the reference strain for fimA genotype V reacted with the primers specific for this genotype. Twenty-one isolates (25.6%) were positive by more than one of the fimA PCR assays; the most frequent combinations were genotypes I, Ib, and II (eight isolates) and genotypes I and II (four isolates). Sequencing of the fimA gene from selected isolates did not support the observed specific fimA genotype combinations, suggesting that the genotyping method used for the major fimbriae in P. gingivalis should be reevaluated.
Insights
The fimA genotyping method for Porphyromonas gingivalis needs reevaluation, as PCR assays showed inconsistent results with genetic sequencing. Genotype II was most common in periodontitis patients.
Area of Science:
- Microbiology
- Genetics
- Periodontal disease
Background:
- Fimbriae are crucial virulence factors in pathogenic bacteria like Porphyromonas gingivalis.
- The fimA gene in P. gingivalis exhibits six distinct genotypes (I, Ib, II, III, IV, V).
- Genotypes II and IV are frequently found in periodontitis patients.
Purpose of the Study:
- To analyze fimA genotype distribution in 82 P. gingivalis isolates from periodontitis patients globally.
- To explore the correlation between fimA genotypes and multilocus sequence typing (MLST) sequence types (STs).
Main Methods:
- Polymerase chain reaction (PCR) with genotype-specific primers was used to amplify the fimA gene.
- Multilocus sequence typing (MLST) was performed to determine the sequence types (STs) of the isolates.
- FimA gene sequencing was conducted on selected isolates.
Main Results:
- A high degree of genetic diversity was observed, with 82 strains assigned to 69 STs.
- FimA genotype II was identified in 34.1% of strains; genotype V was detected only in the reference strain.
- Multiple fimA genotypes were detected in 25.6% of isolates, but sequencing did not support these combinations.
Conclusions:
- The study highlights extensive genetic diversity within P. gingivalis populations.
- Observed discrepancies between PCR-based fimA genotyping and sequencing suggest limitations in the current method.
- Reevaluation of the fimA genotyping methodology for P. gingivalis is recommended.
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