Comprehensive analysis of secondary dental root canal infections : a combination of culture and culture-independent approaches reveals new insights
Persistence of microorganisms or reinfections are the main reasons for failure of root canal therapy. Very few studies to date have included culture-independent methods to assess the microbiota, including non-cultivable microorganisms. The aim of this study was to combine culture methods with culture-independent cloning methods to analyze the microbial flora of root-filled teeth with periradicular lesions. Twenty-one samples from previously root-filled teeth were collected from patients with periradicular lesions. Microorganisms were cultivated, isolated and biochemically identified. In addition, ribosomal DNA of bacteria, fungi and archaea derived from the same samples was amplified and the PCR products were used to construct clone libraries. DNA of selected clones was sequenced and microbial species were identified, comparing the sequences with public databases. Microorganisms were found in 12 samples with culture-dependent and -independent methods combined. The number of bacterial species ranged from 1 to 12 in one sample. The majority of the 26 taxa belonged to the phylum Firmicutes (14 taxa), followed by Actinobacteria, Proteobacteria and Bacteroidetes. One sample was positive for fungi, and archaea could not be detected. The results obtained with both methods differed. The cloning technique detected several as-yet-uncultivated taxa. Using a combination of both methods 13 taxa were detected that had not been found in root-filled teeth so far. Enterococcus faecalis was only detected in two samples using culture methods. Combining the culture-dependent and -independent approaches revealed new candidate endodontic pathogens and a high diversity of the microbial flora in root-filled teeth with periradicular lesions. Both methods yielded differing results, emphasizing the benefit of combined methods for the detection of the actual microbial diversity in apical periodontitis.
Medienart: |
E-Artikel |
---|
Erscheinungsjahr: |
2012 |
---|---|
Erschienen: |
2012 |
Enthalten in: |
Zur Gesamtaufnahme - volume:7 |
---|---|
Enthalten in: |
PloS one - 7(2012), 11 vom: 03., Seite e49576 |
Sprache: |
Englisch |
---|
Beteiligte Personen: |
Anderson, Annette Carola [VerfasserIn] |
---|
Links: |
---|
Themen: |
DNA, Ribosomal |
---|
Anmerkungen: |
Date Completed 07.05.2013 Date Revised 09.04.2022 published: Print-Electronic Citation Status MEDLINE |
---|
doi: |
10.1371/journal.pone.0049576 |
---|
funding: |
|
---|---|
Förderinstitution / Projekttitel: |
|
PPN (Katalog-ID): |
NLM222629819 |
---|
LEADER | 01000naa a22002652 4500 | ||
---|---|---|---|
001 | NLM222629819 | ||
003 | DE-627 | ||
005 | 20231224054719.0 | ||
007 | cr uuu---uuuuu | ||
008 | 231224s2012 xx |||||o 00| ||eng c | ||
024 | 7 | |a 10.1371/journal.pone.0049576 |2 doi | |
028 | 5 | 2 | |a pubmed24n0742.xml |
035 | |a (DE-627)NLM222629819 | ||
035 | |a (NLM)23152922 | ||
040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
041 | |a eng | ||
100 | 1 | |a Anderson, Annette Carola |e verfasserin |4 aut | |
245 | 1 | 0 | |a Comprehensive analysis of secondary dental root canal infections |b a combination of culture and culture-independent approaches reveals new insights |
264 | 1 | |c 2012 | |
336 | |a Text |b txt |2 rdacontent | ||
337 | |a ƒaComputermedien |b c |2 rdamedia | ||
338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
500 | |a Date Completed 07.05.2013 | ||
500 | |a Date Revised 09.04.2022 | ||
500 | |a published: Print-Electronic | ||
500 | |a Citation Status MEDLINE | ||
520 | |a Persistence of microorganisms or reinfections are the main reasons for failure of root canal therapy. Very few studies to date have included culture-independent methods to assess the microbiota, including non-cultivable microorganisms. The aim of this study was to combine culture methods with culture-independent cloning methods to analyze the microbial flora of root-filled teeth with periradicular lesions. Twenty-one samples from previously root-filled teeth were collected from patients with periradicular lesions. Microorganisms were cultivated, isolated and biochemically identified. In addition, ribosomal DNA of bacteria, fungi and archaea derived from the same samples was amplified and the PCR products were used to construct clone libraries. DNA of selected clones was sequenced and microbial species were identified, comparing the sequences with public databases. Microorganisms were found in 12 samples with culture-dependent and -independent methods combined. The number of bacterial species ranged from 1 to 12 in one sample. The majority of the 26 taxa belonged to the phylum Firmicutes (14 taxa), followed by Actinobacteria, Proteobacteria and Bacteroidetes. One sample was positive for fungi, and archaea could not be detected. The results obtained with both methods differed. The cloning technique detected several as-yet-uncultivated taxa. Using a combination of both methods 13 taxa were detected that had not been found in root-filled teeth so far. Enterococcus faecalis was only detected in two samples using culture methods. Combining the culture-dependent and -independent approaches revealed new candidate endodontic pathogens and a high diversity of the microbial flora in root-filled teeth with periradicular lesions. Both methods yielded differing results, emphasizing the benefit of combined methods for the detection of the actual microbial diversity in apical periodontitis | ||
650 | 4 | |a Journal Article | |
650 | 4 | |a Research Support, Non-U.S. Gov't | |
650 | 7 | |a DNA, Ribosomal |2 NLM | |
700 | 1 | |a Hellwig, Elmar |e verfasserin |4 aut | |
700 | 1 | |a Vespermann, Robin |e verfasserin |4 aut | |
700 | 1 | |a Wittmer, Annette |e verfasserin |4 aut | |
700 | 1 | |a Schmid, Michael |e verfasserin |4 aut | |
700 | 1 | |a Karygianni, Lamprini |e verfasserin |4 aut | |
700 | 1 | |a Al-Ahmad, Ali |e verfasserin |4 aut | |
773 | 0 | 8 | |i Enthalten in |t PloS one |d 2006 |g 7(2012), 11 vom: 03., Seite e49576 |w (DE-627)NLM167327399 |x 1932-6203 |7 nnns |
773 | 1 | 8 | |g volume:7 |g year:2012 |g number:11 |g day:03 |g pages:e49576 |
856 | 4 | 0 | |u http://dx.doi.org/10.1371/journal.pone.0049576 |3 Volltext |
912 | |a GBV_USEFLAG_A | ||
912 | |a GBV_NLM | ||
951 | |a AR | ||
952 | |d 7 |j 2012 |e 11 |b 03 |h e49576 |