title: The unique hmuY gene sequence as a specific marker of Porphyromonas gingivalis.
abstract: Porphyromonas gingivalis, a major etiological agent of chronic periodontitis, acquires heme from host hemoproteins using the HmuY hemophore. The aim of this study was to develop a specific P. gingivalis marker based on a hmuY gene sequence. Subgingival samples were collected from 66 patients with chronic periodontitis and 40 healthy subjects and the entire hmuY gene was analyzed in positive samples. Phylogenetic analyses demonstrated that both the amino acid sequence of the HmuY protein and the nucleotide sequence of the hmuY gene are unique among P. gingivalis strains/isolates and show low identity to sequences found in other species (below 50 and 56%, respectively). In agreement with these findings, a set of hmuY gene-based primers and standard/real-time PCR with SYBR Green chemistry allowed us to specifically detect P. gingivalis in patients with chronic periodontitis (77.3%) and healthy subjects (20%), the latter possessing lower number of P. gingivalis cells and total bacterial cells. Isolates from healthy subjects possess the hmuY gene-based nucleotide sequence pattern occurring in W83/W50/A7436 (n = 4), 381/ATCC 33277 (n = 3) or TDC60 (n = 1) strains, whereas those from patients typically have TDC60 (n = 21), W83/W50/A7436 (n = 17) and 381/ATCC 33277 (n = 13) strains. We observed a significant correlation between periodontal index of risk of infectiousness (PIRI) and the presence/absence of P. gingivalis (regardless of the hmuY gene-based sequence pattern of the isolate identified [r = 0.43; P = 0.0002] and considering particular isolate pattern [r = 0.38; P = 0.0012]). In conclusion, we demonstrated that the hmuY gene sequence or its fragments may be used as one of the molecular markers of P. gingivalis.
authors.affiliation: Laboratory of Biochemistry, Faculty of Biotechnology, University of Wrocław, Wrocław, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Gmiterek
authors.forename: Halina
authors.initials: H
authors.lastname: Wójtowicz
authors.forename: Paweł
authors.initials: P
authors.lastname: Mackiewicz
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Radwan-Oczko
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Kantorowicz
authors.forename: Maria
authors.initials: M
authors.lastname: Chomyszyn-Gajewska
authors.forename: Magdalena
authors.initials: M
authors.lastname: Frąszczak
authors.forename: Marcin
authors.initials: M
authors.lastname: Bielecki
authors.forename: Mariusz
authors.initials: M
authors.lastname: Olczak
authors.forename: Teresa
authors.initials: T
authors.lastname: Olczak
mesh_terms: D000328:Adult; D000368:Aged; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016866:Bacteroidaceae Infections / Q000382:microbiology*; D001483:Base Sequence; D015415:Biomarkers / Q000032:analysis; D016022:Case-Control Studies; D055113:Chronic Periodontitis / Q000382:microbiology*; D005260:Female; D006418:Heme / Q000378:metabolism; D006420:Hemeproteins / Q000378:metabolism; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010802:Phylogeny; D016966:Porphyromonas gingivalis / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins; D015415:Biomarkers; D006420:Hemeproteins; D006418:Heme