Detection and quantification of Aggregatibacter actinomycetemcomitans,Porphyromonas gingivalis and Streptococcus oralis in blood samples with different microbiological identification methods: An in vitro study |
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| Affiliation: | 1. Oral Research Laboratory, Faculty of Odontology, University Complutense, Madrid, Spain;2. Department of Special Needs, School of Medicine and Dentistry, Santiago de Compostela University, Santiago de Compostela, Spain;3. Research Laboratory, Department of Clinical Microbiology, Xeral-Cíes Hospital, Vigo, Spain;4. Etiology and Therapy of Periodontal Diseases (ETEP) Research Group, University Complutense, Madrid, Spain;1. University of Texas Southwestern Medical Center, Parkland Hospital, Dallas, TX, USA;2. Director, Section of Oral and Maxillofacial Surgery, Dallas Veterans Affairs Medical Center, Dallas, TX, USA; Faculty, Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center, Dallas, TX, USA;3. Professor, Department of Orthodontics, University of North Carolina School of Dentistry, Chapel Hill, NC, USA;1. Faculty of Dentistry, Federal University of Ceará, Sobral, Ceará, Brazil;2. Department of Oral Diagnosis, Piracicaba Dental School, State University of Campinas, Piracicaba, São Paulo, Brazil;3. Faculty of Pharmacy, Dentistry and Nursing, Federal University of Ceará, Fortaleza, Ceará, Brazil;1. EPICIME-CIC 1407 de Lyon, Inserm, service de pharmacologie clinique, hospices civils de Lyon, 69677 Bron, France;2. UMR 5558, laboratoire de biométrie et biologie évolutive, service de pharmacologie clinique, CNRS, université de Lyon, université Lyon 1, 69622 Villeurbanne, France;3. Réseau d’enquête pédiatrique des produits de santé (RIPPS)-KIDS-France, groupement hospitalier Est, bâtiment « Les Tilleuls », 59, boulevard Pinel, 69677 Bron, France;1. Servicio de Neumología, Hospital Universitario La Princesa, Madrid, España;2. Servicio de Microbiología, Hospital Universitario La Princesa, Madrid, España;1. Division of Oral Radiology, Piracicaba Dental School, University of Campinas, Department of Oral Diagnosis, Limeira Avenue, 901, PO Box 52, Piracicaba, SP, Brazil;2. Division of Pharmacology, Anesthesiology and Therapeutic, Piracicaba Dental School, University of Campinas, Department of Physiological Sciences, Limeira Avenue, 901, PO Box 52, Piracicaba, SP, Brazil;3. Division of Oral Radiology, Federal University of Rio Grande do Sul, Department of Surgery and Orthopedics, Porto Alegre, Ramiro Barcelos Avenue, 2492, Porto Alegre, RS, Brazil;4. Department of Oral and Maxillofacial Radiology, Academic Centre for Dentistry Amsterdam (ACTA), 4N-51 Gustav Mahlerlaan 3004, 1081 LA, Amsterdam, The Netherlands |
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| Abstract: | BackgroundCulture-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.ObjectiveTo 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 methodsBlood 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.ResultsDAC, 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.ConclusionsDAC 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. |
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| Keywords: | Bacteremia Quantitative PCR Culture Periodontal |
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