Quantitative reverse transcription polymerase chain reaction analysis of Porphyromonas gingivalis gene expression in vivo

Charles E. Shelburne, Raymond M. Gleason, Gregory R Germaine, Larry F. Wolff, Brian H. Mullally, Wilson A. Coulter, Dennis E. Lopatin

Research output: Contribution to journalArticlepeer-review

27 Citations (Scopus)


An etiological relationship between periodontitis, a significant oral health problem, and the anaerobe Porphyromonas gingivalis may be related to the expression of a variety of putative virulence factors. The objective of the experiments described here was to develop a quantitative reverse transcription polymerase chain reaction (QRT-PCR) method to examine P. gingivalis gene expression in human dental plaque from periodontitis subjects. PCR primers and probes for six target genes representing putative virulence factors were chosen and evaluated in vitro for specificity. A potential cross-reactivity level of only 10 copies/107 whole genomic equivalents was occasionally observed with non-P. gingivalis microbes. P. gingivalis cells stressed in vitro by a 5 °C temperature increase showed a rapid rise in the mRNA associated with the molecular chaperons (htpG, dnaK, groEL), SOD (sodA) and gingipain (rgp-1) genes. We examined the stability of bacterial RNA in plaque specimens and found no significant difference in the amount of RNA obtained before or after storage 3 months in a stabilizing buffer (p=0.786, t-test). Sixty-five percent of plaque samples obtained from two clinical locations contained P. gingivalis; there was a mean level of gene expression (fold increase) for all samples tested for groEL, dnaK, htpG, sodA, PG1431 and rgp-1 of 0.84±2.03 to 7.85±10.0. ANOVA showed that the levels of stress gene transcription for dnaK and htpG were significantly elevated (p
Original languageEnglish
Pages (from-to)147-156
Number of pages10
JournalJournal of Microbiological Methods
Issue number2
Early online date26 Nov 2001
Publication statusPublished - Apr 2002

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Microbiology


Dive into the research topics of 'Quantitative reverse transcription polymerase chain reaction analysis of Porphyromonas gingivalis gene expression in vivo'. Together they form a unique fingerprint.

Cite this