Cargando…

Application of a target enrichment-based next-generation sequencing protocol for identification and sequence-based prediction of pneumococcal serotypes

BACKGROUND: The use of whole-genome sequencing in microbiology at a diagnostic level, although feasible, is still limited by the expenses associated and by the complex bioinformatics pipelines in data analyses. We describe the use of target enrichment-based next-generation sequencing for pneumococca...

Descripción completa

Detalles Bibliográficos
Autores principales: Liyanapathirana, Veranja, Ang, Irene, Tsang, Dominic, Fung, Kitty, Ng, Tak Keung, Zhou, Haokui, Ip, Margaret
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3984734/
https://www.ncbi.nlm.nih.gov/pubmed/24612771
http://dx.doi.org/10.1186/1471-2180-14-60
_version_ 1782311478824534016
author Liyanapathirana, Veranja
Ang, Irene
Tsang, Dominic
Fung, Kitty
Ng, Tak Keung
Zhou, Haokui
Ip, Margaret
author_facet Liyanapathirana, Veranja
Ang, Irene
Tsang, Dominic
Fung, Kitty
Ng, Tak Keung
Zhou, Haokui
Ip, Margaret
author_sort Liyanapathirana, Veranja
collection PubMed
description BACKGROUND: The use of whole-genome sequencing in microbiology at a diagnostic level, although feasible, is still limited by the expenses associated and by the complex bioinformatics pipelines in data analyses. We describe the use of target enrichment-based next-generation sequencing for pneumococcal identification and serotyping as applied to the polysaccharide 23 valent vaccine serotypes as an affordable alternative to whole genome sequencing. RESULTS: Correct identification of Streptococcus pneumoniae and prediction of common vaccine serotypes: 12 to serotype level and the rest to serogroup levels were achieved for all serotypes with >500 reads mapped against serotypes sequences. A proportion-based criterion also enabled the identification of two serotypes present in the same sample, thus indicating the possibility of using this method in detecting co-colonizing serotypes. The results obtained were comparable to or an improvement on the currently existing molecular serotyping methods for S. pneumoniae in relation to the polysaccharide vaccine serotypes. CONCLUSION: We propose that this method has the potential to become an affordable and adaptable alternative to whole-genome sequencing for pneumococcal identification and serotyping.
format Online
Article
Text
id pubmed-3984734
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-39847342014-04-14 Application of a target enrichment-based next-generation sequencing protocol for identification and sequence-based prediction of pneumococcal serotypes Liyanapathirana, Veranja Ang, Irene Tsang, Dominic Fung, Kitty Ng, Tak Keung Zhou, Haokui Ip, Margaret BMC Microbiol Research Article BACKGROUND: The use of whole-genome sequencing in microbiology at a diagnostic level, although feasible, is still limited by the expenses associated and by the complex bioinformatics pipelines in data analyses. We describe the use of target enrichment-based next-generation sequencing for pneumococcal identification and serotyping as applied to the polysaccharide 23 valent vaccine serotypes as an affordable alternative to whole genome sequencing. RESULTS: Correct identification of Streptococcus pneumoniae and prediction of common vaccine serotypes: 12 to serotype level and the rest to serogroup levels were achieved for all serotypes with >500 reads mapped against serotypes sequences. A proportion-based criterion also enabled the identification of two serotypes present in the same sample, thus indicating the possibility of using this method in detecting co-colonizing serotypes. The results obtained were comparable to or an improvement on the currently existing molecular serotyping methods for S. pneumoniae in relation to the polysaccharide vaccine serotypes. CONCLUSION: We propose that this method has the potential to become an affordable and adaptable alternative to whole-genome sequencing for pneumococcal identification and serotyping. BioMed Central 2014-03-10 /pmc/articles/PMC3984734/ /pubmed/24612771 http://dx.doi.org/10.1186/1471-2180-14-60 Text en Copyright © 2014 Liyanapathirana et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Liyanapathirana, Veranja
Ang, Irene
Tsang, Dominic
Fung, Kitty
Ng, Tak Keung
Zhou, Haokui
Ip, Margaret
Application of a target enrichment-based next-generation sequencing protocol for identification and sequence-based prediction of pneumococcal serotypes
title Application of a target enrichment-based next-generation sequencing protocol for identification and sequence-based prediction of pneumococcal serotypes
title_full Application of a target enrichment-based next-generation sequencing protocol for identification and sequence-based prediction of pneumococcal serotypes
title_fullStr Application of a target enrichment-based next-generation sequencing protocol for identification and sequence-based prediction of pneumococcal serotypes
title_full_unstemmed Application of a target enrichment-based next-generation sequencing protocol for identification and sequence-based prediction of pneumococcal serotypes
title_short Application of a target enrichment-based next-generation sequencing protocol for identification and sequence-based prediction of pneumococcal serotypes
title_sort application of a target enrichment-based next-generation sequencing protocol for identification and sequence-based prediction of pneumococcal serotypes
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3984734/
https://www.ncbi.nlm.nih.gov/pubmed/24612771
http://dx.doi.org/10.1186/1471-2180-14-60
work_keys_str_mv AT liyanapathiranaveranja applicationofatargetenrichmentbasednextgenerationsequencingprotocolforidentificationandsequencebasedpredictionofpneumococcalserotypes
AT angirene applicationofatargetenrichmentbasednextgenerationsequencingprotocolforidentificationandsequencebasedpredictionofpneumococcalserotypes
AT tsangdominic applicationofatargetenrichmentbasednextgenerationsequencingprotocolforidentificationandsequencebasedpredictionofpneumococcalserotypes
AT fungkitty applicationofatargetenrichmentbasednextgenerationsequencingprotocolforidentificationandsequencebasedpredictionofpneumococcalserotypes
AT ngtakkeung applicationofatargetenrichmentbasednextgenerationsequencingprotocolforidentificationandsequencebasedpredictionofpneumococcalserotypes
AT zhouhaokui applicationofatargetenrichmentbasednextgenerationsequencingprotocolforidentificationandsequencebasedpredictionofpneumococcalserotypes
AT ipmargaret applicationofatargetenrichmentbasednextgenerationsequencingprotocolforidentificationandsequencebasedpredictionofpneumococcalserotypes