Cargando…
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...
Autores principales: | , , , |
---|---|
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 |
_version_ | 1782253282403549184 |
---|---|
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. |
format | Online Article Text |
id | pubmed-3524157 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Landes Bioscience |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT borekannal typingofstreptococcuspyogenesstrainsusingthephageprofilingmethod AT obszanskakatarzyna typingofstreptococcuspyogenesstrainsusingthephageprofilingmethod AT hryniewiczwaleria typingofstreptococcuspyogenesstrainsusingthephageprofilingmethod AT sitkiewiczizabela typingofstreptococcuspyogenesstrainsusingthephageprofilingmethod |