The unique hmuY gene sequence as a specific marker of Porphyromonas gingivalis

• Published in: PloS one 2013-07, Vol.8 (7), p.e67719-e67719
• Main Authors: Gmiterek, Anna, Wójtowicz, Halina, Mackiewicz, Paweł, Radwan-Oczko, Małgorzata, Kantorowicz, Małgorzata, Chomyszyn-Gajewska, Maria, Frąszczak, Magdalena, Bielecki, Marcin, Olczak, Mariusz, Olczak, Teresa
• Format: Article
• Language: English
• Subjects: Adult; Aged; Amino acid sequence; Anaerobic bacteria; Antibiotics; Bacteria; Bacterial infections; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteroidaceae Infections - microbiology; Bacteroides; Bacteroides gingivalis; Base Sequence; Biochemistry; Biological markers; Biology; Biomarkers - analysis; Biotechnology; Case-Control Studies; Chronic Periodontitis - microbiology; Deoxyribonucleic acid; Disease; DNA; Etiology; Female; Genes; Genetic aspects; Genomes; Genomics; Green chemistry; Green development; Gum disease; Health risks; Heme; Heme - metabolism; Hemeproteins - metabolism; HmuY protein; Humans; Hybridization; Infections; Laboratories; Male; Medicine; Microbiota; Middle Aged; Molecular Sequence Data; Nucleotide sequence; Pathogens; Patients; Periodontitis; Phylogeny; Porphyromonas gingivalis; Porphyromonas gingivalis - classification; Porphyromonas gingivalis - genetics; Porphyromonas gingivalis - isolation & purification; Primers; RNA polymerase; Sequence Alignment; Strains (organisms); Teeth
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0067719

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.