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RAPD-PCR-Based Fingerprinting Method as a Tool for Epidemiological Analysis of Trueperella pyogenes Infections

In this study, a Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) method for genetic typing of Trueperella pyogenes, an opportunistic bacterial pathogen, was designed. The method optimization was performed for 37 clinical T. pyogenes strains isolated from various infections in d...

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Autores principales: Stefańska, Ilona, Kwiecień, Ewelina, Górzyńska, Małgorzata, Sałamaszyńska-Guz, Agnieszka, Rzewuska, Magdalena
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9147813/
https://www.ncbi.nlm.nih.gov/pubmed/35631083
http://dx.doi.org/10.3390/pathogens11050562
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author Stefańska, Ilona
Kwiecień, Ewelina
Górzyńska, Małgorzata
Sałamaszyńska-Guz, Agnieszka
Rzewuska, Magdalena
author_facet Stefańska, Ilona
Kwiecień, Ewelina
Górzyńska, Małgorzata
Sałamaszyńska-Guz, Agnieszka
Rzewuska, Magdalena
author_sort Stefańska, Ilona
collection PubMed
description In this study, a Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) method for genetic typing of Trueperella pyogenes, an opportunistic bacterial pathogen, was designed. The method optimization was performed for 37 clinical T. pyogenes strains isolated from various infections in different animal species. Optimal conditions for reliable and reproducible DNA fingerprinting were determined according to the modified Taguchi method. The developed method was assessed regarding its typeability, reproducibility, and discriminatory power using the Hunter’s and Gatsons’ index of discrimination. A high degree of genetic diversity was shown between the studied strains, which represented 31 genotypes. The generated RAPD profiles were relatively complex and simultaneously easy to interpret due to the wide size range of amplicons. The discriminatory index of the designed method was sufficiently high; thus, only strains epidemiologically related displayed identical RAPD genotypes. In conclusion, the DNA fingerprinting of T. pyogenes by the developed RAPD-PCR method is a reliable typing tool that may allow a better understanding of the epidemiology as well as pathogenesis of infections caused by this pathogen.
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spelling pubmed-91478132022-05-29 RAPD-PCR-Based Fingerprinting Method as a Tool for Epidemiological Analysis of Trueperella pyogenes Infections Stefańska, Ilona Kwiecień, Ewelina Górzyńska, Małgorzata Sałamaszyńska-Guz, Agnieszka Rzewuska, Magdalena Pathogens Article In this study, a Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) method for genetic typing of Trueperella pyogenes, an opportunistic bacterial pathogen, was designed. The method optimization was performed for 37 clinical T. pyogenes strains isolated from various infections in different animal species. Optimal conditions for reliable and reproducible DNA fingerprinting were determined according to the modified Taguchi method. The developed method was assessed regarding its typeability, reproducibility, and discriminatory power using the Hunter’s and Gatsons’ index of discrimination. A high degree of genetic diversity was shown between the studied strains, which represented 31 genotypes. The generated RAPD profiles were relatively complex and simultaneously easy to interpret due to the wide size range of amplicons. The discriminatory index of the designed method was sufficiently high; thus, only strains epidemiologically related displayed identical RAPD genotypes. In conclusion, the DNA fingerprinting of T. pyogenes by the developed RAPD-PCR method is a reliable typing tool that may allow a better understanding of the epidemiology as well as pathogenesis of infections caused by this pathogen. MDPI 2022-05-10 /pmc/articles/PMC9147813/ /pubmed/35631083 http://dx.doi.org/10.3390/pathogens11050562 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Stefańska, Ilona
Kwiecień, Ewelina
Górzyńska, Małgorzata
Sałamaszyńska-Guz, Agnieszka
Rzewuska, Magdalena
RAPD-PCR-Based Fingerprinting Method as a Tool for Epidemiological Analysis of Trueperella pyogenes Infections
title RAPD-PCR-Based Fingerprinting Method as a Tool for Epidemiological Analysis of Trueperella pyogenes Infections
title_full RAPD-PCR-Based Fingerprinting Method as a Tool for Epidemiological Analysis of Trueperella pyogenes Infections
title_fullStr RAPD-PCR-Based Fingerprinting Method as a Tool for Epidemiological Analysis of Trueperella pyogenes Infections
title_full_unstemmed RAPD-PCR-Based Fingerprinting Method as a Tool for Epidemiological Analysis of Trueperella pyogenes Infections
title_short RAPD-PCR-Based Fingerprinting Method as a Tool for Epidemiological Analysis of Trueperella pyogenes Infections
title_sort rapd-pcr-based fingerprinting method as a tool for epidemiological analysis of trueperella pyogenes infections
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9147813/
https://www.ncbi.nlm.nih.gov/pubmed/35631083
http://dx.doi.org/10.3390/pathogens11050562
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