Detection and quantification of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Streptococcus oralis in blood samples with different microbiological identification methods: An in vitro study

Arch Oral Biol. 2017 Feb:74:55-62. doi: 10.1016/j.archoralbio.2016.11.007. Epub 2016 Nov 9.

Abstract

Background: Culture-based methods (culture broth bottles or lysis methods) have been the standard for detecting bacteremia. More recently, quantitative polymerase chain reaction (qPCR) was proposed as a more sensitive and specific test although none of them has been validated for the identification of periodontal pathogens (fastidious growing bacteria) in blood samples.

Objective: To compare the ability to detect and quantify Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Streptococcus oralis (alone or in combination) in blood samples with three culture techniques [direct anaerobic culturing (DAC), haemo-culture (BACTEC), and lysis-centrifugation (LC)] and a non-culture dependent approach (qPCR) in an in vitro study.

Material and methods: Blood samples from 12 periodontally healthy volunteers were contaminated with three concentrations [104,102 and 101 colony forming units (CFU)/mL] of A. actinomycetemcomitans, P. gingivalis and S. oralis, alone or in combination. Samples were analysed by DAC, BACTEC, LC and qPCR. Sensitivity, specificity, predictive values, kappa index and Lińs correlation coefficients were calculated.

Results: DAC, LC and qPCR were able to detect the three target species at all concentrations. An excellent concordance (correlation coefficient r: 0.92-1) was observed between DAC and the reference standard (sensitivity raging 93.33-100% and specificity 88.89-100%) values. BACTEC was not able to identify P. gingivalis in any of the performed experiments. qPCR provided false negative results for S.oralis.

Conclusions: DAC showed the best results for the proper identification and quantification of A. actinomycetemcomitans, P. gingivalis and S. oralis, alone or in combination, in blood samples.

Keywords: Aggregatibacter actinomycetemitans; Bacteremia; Culture; Periodontal; Porphyromonas gingivalis; Quantitative PCR.

MeSH terms

  • Aggregatibacter actinomycetemcomitans / genetics
  • Aggregatibacter actinomycetemcomitans / isolation & purification*
  • Bacteremia / diagnosis
  • Bacteremia / microbiology
  • Bacterial Load
  • Bacteriological Techniques / instrumentation
  • Bacteriological Techniques / methods*
  • Blood / microbiology*
  • Colony Count, Microbial
  • Culture Techniques / methods
  • DNA, Bacterial / analysis
  • DNA, Bacterial / isolation & purification
  • Genes, Bacterial / genetics
  • Humans
  • Periodontal Diseases / diagnosis
  • Periodontal Diseases / microbiology
  • Polymerase Chain Reaction / methods
  • Porphyromonas gingivalis / genetics
  • Porphyromonas gingivalis / isolation & purification*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Streptococcus oralis / genetics
  • Streptococcus oralis / isolation & purification*

Substances

  • DNA, Bacterial