Cargando…

Advantages and Limitations of Ribosomal RNA PCR and DNA Sequencing for Identification of Bacteria in Cardiac Valves of Danish Patients

Studies on the value of culture-independent molecular identification of bacteria in cardiac valves are mostly restricted to comparing agreement of identification to what is obtained by culture to the number of identified bacteria in culture-negative cases. However, evaluation of the usefulness of di...

Descripción completa

Detalles Bibliográficos
Autores principales: Kemp, Michael, Bangsborg, Jette, Kjerulf, Anne, Schmidt, Thomas Andersen, Christensen, John, Irmukhamedov, Akhmadjon 6, Bruun, Niels Eske, Dargis, Rimtas, Andresen, Keld, Christensen, Jens Jørgen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Bentham Open 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3883160/
https://www.ncbi.nlm.nih.gov/pubmed/24403979
http://dx.doi.org/10.2174/1874285801307010146
_version_ 1782298408149581824
author Kemp, Michael
Bangsborg, Jette
Kjerulf, Anne
Schmidt, Thomas Andersen
Christensen, John
Irmukhamedov, Akhmadjon 6
Bruun, Niels Eske
Dargis, Rimtas
Andresen, Keld
Christensen, Jens Jørgen
author_facet Kemp, Michael
Bangsborg, Jette
Kjerulf, Anne
Schmidt, Thomas Andersen
Christensen, John
Irmukhamedov, Akhmadjon 6
Bruun, Niels Eske
Dargis, Rimtas
Andresen, Keld
Christensen, Jens Jørgen
author_sort Kemp, Michael
collection PubMed
description Studies on the value of culture-independent molecular identification of bacteria in cardiac valves are mostly restricted to comparing agreement of identification to what is obtained by culture to the number of identified bacteria in culture-negative cases. However, evaluation of the usefulness of direct molecular identification should also address weaknesses, their relevance in the given setting, and possible improvements. In this study cardiac valves from 56 Danish patients referred for surgery for infective endocarditis were analysed by microscopy and culture as well as by PCR targeting part of the bacterial 16S rRNA gene followed by DNA sequencing of the PCR product. PCR and DNA sequencing identified significant bacteria in 49 samples from 43 patients, including five out of 13 culture-negative cases. No rare, exotic, or intracellular bacteria were identified. There was a general agreement between bacterial identity obtained by ribosomal PCR and DNA sequencing from the valves and bacterial isolates from blood culture. However, DNA sequencing of the 16S rRNA gene did not discriminate well among non-haemolytic streptococci, especially within the Streptococcus mitis group. Ribosomal PCR with subsequent DNA sequencing is an efficient and reliable method of identifying the cause of IE, but exact species identification of some of the most common causes, i.e. non-haemolytic streptococci, may be improved with other molecular methods.
format Online
Article
Text
id pubmed-3883160
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Bentham Open
record_format MEDLINE/PubMed
spelling pubmed-38831602014-01-08 Advantages and Limitations of Ribosomal RNA PCR and DNA Sequencing for Identification of Bacteria in Cardiac Valves of Danish Patients Kemp, Michael Bangsborg, Jette Kjerulf, Anne Schmidt, Thomas Andersen Christensen, John Irmukhamedov, Akhmadjon 6 Bruun, Niels Eske Dargis, Rimtas Andresen, Keld Christensen, Jens Jørgen Open Microbiol J Article Studies on the value of culture-independent molecular identification of bacteria in cardiac valves are mostly restricted to comparing agreement of identification to what is obtained by culture to the number of identified bacteria in culture-negative cases. However, evaluation of the usefulness of direct molecular identification should also address weaknesses, their relevance in the given setting, and possible improvements. In this study cardiac valves from 56 Danish patients referred for surgery for infective endocarditis were analysed by microscopy and culture as well as by PCR targeting part of the bacterial 16S rRNA gene followed by DNA sequencing of the PCR product. PCR and DNA sequencing identified significant bacteria in 49 samples from 43 patients, including five out of 13 culture-negative cases. No rare, exotic, or intracellular bacteria were identified. There was a general agreement between bacterial identity obtained by ribosomal PCR and DNA sequencing from the valves and bacterial isolates from blood culture. However, DNA sequencing of the 16S rRNA gene did not discriminate well among non-haemolytic streptococci, especially within the Streptococcus mitis group. Ribosomal PCR with subsequent DNA sequencing is an efficient and reliable method of identifying the cause of IE, but exact species identification of some of the most common causes, i.e. non-haemolytic streptococci, may be improved with other molecular methods. Bentham Open 2013-12-27 /pmc/articles/PMC3883160/ /pubmed/24403979 http://dx.doi.org/10.2174/1874285801307010146 Text en © Kemp et al.; Licensee Bentham Open. http://creativecommons.org/licenses/by-nc/3.0/ This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.
spellingShingle Article
Kemp, Michael
Bangsborg, Jette
Kjerulf, Anne
Schmidt, Thomas Andersen
Christensen, John
Irmukhamedov, Akhmadjon 6
Bruun, Niels Eske
Dargis, Rimtas
Andresen, Keld
Christensen, Jens Jørgen
Advantages and Limitations of Ribosomal RNA PCR and DNA Sequencing for Identification of Bacteria in Cardiac Valves of Danish Patients
title Advantages and Limitations of Ribosomal RNA PCR and DNA Sequencing for Identification of Bacteria in Cardiac Valves of Danish Patients
title_full Advantages and Limitations of Ribosomal RNA PCR and DNA Sequencing for Identification of Bacteria in Cardiac Valves of Danish Patients
title_fullStr Advantages and Limitations of Ribosomal RNA PCR and DNA Sequencing for Identification of Bacteria in Cardiac Valves of Danish Patients
title_full_unstemmed Advantages and Limitations of Ribosomal RNA PCR and DNA Sequencing for Identification of Bacteria in Cardiac Valves of Danish Patients
title_short Advantages and Limitations of Ribosomal RNA PCR and DNA Sequencing for Identification of Bacteria in Cardiac Valves of Danish Patients
title_sort advantages and limitations of ribosomal rna pcr and dna sequencing for identification of bacteria in cardiac valves of danish patients
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3883160/
https://www.ncbi.nlm.nih.gov/pubmed/24403979
http://dx.doi.org/10.2174/1874285801307010146
work_keys_str_mv AT kempmichael advantagesandlimitationsofribosomalrnapcranddnasequencingforidentificationofbacteriaincardiacvalvesofdanishpatients
AT bangsborgjette advantagesandlimitationsofribosomalrnapcranddnasequencingforidentificationofbacteriaincardiacvalvesofdanishpatients
AT kjerulfanne advantagesandlimitationsofribosomalrnapcranddnasequencingforidentificationofbacteriaincardiacvalvesofdanishpatients
AT schmidtthomasandersen advantagesandlimitationsofribosomalrnapcranddnasequencingforidentificationofbacteriaincardiacvalvesofdanishpatients
AT christensenjohn advantagesandlimitationsofribosomalrnapcranddnasequencingforidentificationofbacteriaincardiacvalvesofdanishpatients
AT irmukhamedovakhmadjon6 advantagesandlimitationsofribosomalrnapcranddnasequencingforidentificationofbacteriaincardiacvalvesofdanishpatients
AT bruunnielseske advantagesandlimitationsofribosomalrnapcranddnasequencingforidentificationofbacteriaincardiacvalvesofdanishpatients
AT dargisrimtas advantagesandlimitationsofribosomalrnapcranddnasequencingforidentificationofbacteriaincardiacvalvesofdanishpatients
AT andresenkeld advantagesandlimitationsofribosomalrnapcranddnasequencingforidentificationofbacteriaincardiacvalvesofdanishpatients
AT christensenjensjørgen advantagesandlimitationsofribosomalrnapcranddnasequencingforidentificationofbacteriaincardiacvalvesofdanishpatients