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Typing of Streptococcus pyogenes strains using the phage profiling method

We recently developed a method that allows fast differentiation between Streptococcus pyogenes (GAS) strains. The method named phage profiling (PP) is based on a simple assumption that a regular PCR reaction with Taq polymerase and relatively short elongation time is not able to yield long DNA fragm...

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Autores principales: Borek, Anna L., Obszańska, Katarzyna, Hryniewicz, Waleria, Sitkiewicz, Izabela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Landes Bioscience 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3524157/
https://www.ncbi.nlm.nih.gov/pubmed/23076280
http://dx.doi.org/10.4161/viru.21887
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author Borek, Anna L.
Obszańska, Katarzyna
Hryniewicz, Waleria
Sitkiewicz, Izabela
author_facet Borek, Anna L.
Obszańska, Katarzyna
Hryniewicz, Waleria
Sitkiewicz, Izabela
author_sort Borek, Anna L.
collection PubMed
description We recently developed a method that allows fast differentiation between Streptococcus pyogenes (GAS) strains. The method named phage profiling (PP) is based on a simple assumption that a regular PCR reaction with Taq polymerase and relatively short elongation time is not able to yield long DNA fragment, such as ~40–50 kb integrated prophage. Only fragments without any integrated DNA or short fragments inserted between integration sites can be efficiently amplified. We designed primers that anneal upstream and downstream prophage integration sites, so in simple PCR reaction we can test if any additional DNA is integrated into particular site. Profiling of integrated elements can be used as rapid, high resolution typing method, with the resolution as high as PFGE and is excellent predictor of PFGE type.
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spelling pubmed-35241572012-12-18 Typing of Streptococcus pyogenes strains using the phage profiling method Borek, Anna L. Obszańska, Katarzyna Hryniewicz, Waleria Sitkiewicz, Izabela Virulence Protocol We recently developed a method that allows fast differentiation between Streptococcus pyogenes (GAS) strains. The method named phage profiling (PP) is based on a simple assumption that a regular PCR reaction with Taq polymerase and relatively short elongation time is not able to yield long DNA fragment, such as ~40–50 kb integrated prophage. Only fragments without any integrated DNA or short fragments inserted between integration sites can be efficiently amplified. We designed primers that anneal upstream and downstream prophage integration sites, so in simple PCR reaction we can test if any additional DNA is integrated into particular site. Profiling of integrated elements can be used as rapid, high resolution typing method, with the resolution as high as PFGE and is excellent predictor of PFGE type. Landes Bioscience 2012-10-01 /pmc/articles/PMC3524157/ /pubmed/23076280 http://dx.doi.org/10.4161/viru.21887 Text en Copyright © 2012 Landes Bioscience http://creativecommons.org/licenses/by-nc/3.0/ This is an open-access article licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License. The article may be redistributed, reproduced, and reused for non-commercial purposes, provided the original source is properly cited.
spellingShingle Protocol
Borek, Anna L.
Obszańska, Katarzyna
Hryniewicz, Waleria
Sitkiewicz, Izabela
Typing of Streptococcus pyogenes strains using the phage profiling method
title Typing of Streptococcus pyogenes strains using the phage profiling method
title_full Typing of Streptococcus pyogenes strains using the phage profiling method
title_fullStr Typing of Streptococcus pyogenes strains using the phage profiling method
title_full_unstemmed Typing of Streptococcus pyogenes strains using the phage profiling method
title_short Typing of Streptococcus pyogenes strains using the phage profiling method
title_sort typing of streptococcus pyogenes strains using the phage profiling method
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3524157/
https://www.ncbi.nlm.nih.gov/pubmed/23076280
http://dx.doi.org/10.4161/viru.21887
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