Rapid Multiplex Real-Time PCR Method for the Detection and Quantification of Selected Cariogenic and Periodontal Bacteria

Dental caries and periodontal diseases are associated with a shift from symbiotic microbiota to dysbiosis. The aim of our study was to develop a rapid, sensitive, and economical method for the identification and quantification of selected cariogenic and periodontal oral bacteria. Original protocols...

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Main Authors: Jan Lochman, Martina Zapletalova, Hana Poskerova, Lydie Izakovicova Holla, Petra Borilova Linhartova
Format: Article
Language:English
Published: MDPI AG 2019-12-01
Series:Diagnostics
Subjects:
Online Access:https://www.mdpi.com/2075-4418/10/1/8
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author Jan Lochman
Martina Zapletalova
Hana Poskerova
Lydie Izakovicova Holla
Petra Borilova Linhartova
author_facet Jan Lochman
Martina Zapletalova
Hana Poskerova
Lydie Izakovicova Holla
Petra Borilova Linhartova
author_sort Jan Lochman
collection DOAJ
description Dental caries and periodontal diseases are associated with a shift from symbiotic microbiota to dysbiosis. The aim of our study was to develop a rapid, sensitive, and economical method for the identification and quantification of selected cariogenic and periodontal oral bacteria. Original protocols were designed for three real-time multiplex PCR assays to detect and quantify the ratio of 10 bacterial species associated with dental caries (&#8220;cariogenic&#8221; complex) or periodontal diseases (red complex, orange complex, and <i>Aggregatibacter actinomycetemcomitans</i>). A total number of 60 samples from 30 children aged 2&#8722;6 years with severe early childhood caries and gingivitis were tested. In multiplex assays, the quantification of total bacterial (TB) content for cariogenic bacteria and red complex to eliminate differences in quantities caused by specimen collection was included. The mean counts for the TB load and that of ten evaluated specimens corresponded to previously published results. We found a significant difference between the microbial compositions obtained from the area of control and the affected teeth (<i>p</i> &lt; 0.05). Based on this comprehensive microbiological examination, the risk of dental caries or periodontal inflammation may be determined. The test could also be used as a tool for behavioral intervention and thus prevention of the above-mentioned diseases.
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spelling doaj.art-76c786ec64384fe09d10b18222e37d512022-12-22T04:04:00ZengMDPI AGDiagnostics2075-44182019-12-01101810.3390/diagnostics10010008diagnostics10010008Rapid Multiplex Real-Time PCR Method for the Detection and Quantification of Selected Cariogenic and Periodontal BacteriaJan Lochman0Martina Zapletalova1Hana Poskerova2Lydie Izakovicova Holla3Petra Borilova Linhartova4Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 611 37 Brno, Czech RepublicDepartment of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 611 37 Brno, Czech RepublicClinic of Stomatology, Institution Shared with St. Anne’s Faculty Hospital, Faculty of Medicine, Masaryk University, Pekarska 53, 656 91 Brno, Czech RepublicClinic of Stomatology, Institution Shared with St. Anne’s Faculty Hospital, Faculty of Medicine, Masaryk University, Pekarska 53, 656 91 Brno, Czech RepublicDepartment of Pathophysiology, Faculty of Medicine, Masaryk University, Kamenice 5, 625 00 Brno, Czech RepublicDental caries and periodontal diseases are associated with a shift from symbiotic microbiota to dysbiosis. The aim of our study was to develop a rapid, sensitive, and economical method for the identification and quantification of selected cariogenic and periodontal oral bacteria. Original protocols were designed for three real-time multiplex PCR assays to detect and quantify the ratio of 10 bacterial species associated with dental caries (&#8220;cariogenic&#8221; complex) or periodontal diseases (red complex, orange complex, and <i>Aggregatibacter actinomycetemcomitans</i>). A total number of 60 samples from 30 children aged 2&#8722;6 years with severe early childhood caries and gingivitis were tested. In multiplex assays, the quantification of total bacterial (TB) content for cariogenic bacteria and red complex to eliminate differences in quantities caused by specimen collection was included. The mean counts for the TB load and that of ten evaluated specimens corresponded to previously published results. We found a significant difference between the microbial compositions obtained from the area of control and the affected teeth (<i>p</i> &lt; 0.05). Based on this comprehensive microbiological examination, the risk of dental caries or periodontal inflammation may be determined. The test could also be used as a tool for behavioral intervention and thus prevention of the above-mentioned diseases.https://www.mdpi.com/2075-4418/10/1/8dental cariesperiodontitisdysbiosisoral microbiomerisk factors determination
spellingShingle Jan Lochman
Martina Zapletalova
Hana Poskerova
Lydie Izakovicova Holla
Petra Borilova Linhartova
Rapid Multiplex Real-Time PCR Method for the Detection and Quantification of Selected Cariogenic and Periodontal Bacteria
Diagnostics
dental caries
periodontitis
dysbiosis
oral microbiome
risk factors determination
title Rapid Multiplex Real-Time PCR Method for the Detection and Quantification of Selected Cariogenic and Periodontal Bacteria
title_full Rapid Multiplex Real-Time PCR Method for the Detection and Quantification of Selected Cariogenic and Periodontal Bacteria
title_fullStr Rapid Multiplex Real-Time PCR Method for the Detection and Quantification of Selected Cariogenic and Periodontal Bacteria
title_full_unstemmed Rapid Multiplex Real-Time PCR Method for the Detection and Quantification of Selected Cariogenic and Periodontal Bacteria
title_short Rapid Multiplex Real-Time PCR Method for the Detection and Quantification of Selected Cariogenic and Periodontal Bacteria
title_sort rapid multiplex real time pcr method for the detection and quantification of selected cariogenic and periodontal bacteria
topic dental caries
periodontitis
dysbiosis
oral microbiome
risk factors determination
url https://www.mdpi.com/2075-4418/10/1/8
work_keys_str_mv AT janlochman rapidmultiplexrealtimepcrmethodforthedetectionandquantificationofselectedcariogenicandperiodontalbacteria
AT martinazapletalova rapidmultiplexrealtimepcrmethodforthedetectionandquantificationofselectedcariogenicandperiodontalbacteria
AT hanaposkerova rapidmultiplexrealtimepcrmethodforthedetectionandquantificationofselectedcariogenicandperiodontalbacteria
AT lydieizakovicovaholla rapidmultiplexrealtimepcrmethodforthedetectionandquantificationofselectedcariogenicandperiodontalbacteria
AT petraborilovalinhartova rapidmultiplexrealtimepcrmethodforthedetectionandquantificationofselectedcariogenicandperiodontalbacteria