A population-level strain genotyping method to study pathogen strain dynamics in human infections
A hallmark of chronic bacterial infections is the long-term persistence of 1 or more pathogen species at the compromised site. Repeated detection of the same bacterial species can suggest that a single strain or lineage is continually present. However, infection with multiple strains of a given spec...
Ausführliche Beschreibung
Autor*in: |
Sarah J. Morgan [verfasserIn] Samantha L. Durfey [verfasserIn] Sumedha Ravishankar [verfasserIn] Peter Jorth [verfasserIn] Wendy Ni [verfasserIn] Duncan T. Skerrett [verfasserIn] Moira L. Aitken [verfasserIn] Edward F. McKone [verfasserIn] Stephen J. Salipante [verfasserIn] Matthew C. Radey [verfasserIn] Pradeep K. Singh [verfasserIn] |
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E-Artikel |
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Englisch |
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2021 |
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In: JCI Insight - American Society for Clinical investigation, 2020, 6(2021), 24 |
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Übergeordnetes Werk: |
volume:6 ; year:2021 ; number:24 |
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Katalog-ID: |
DOAJ020422164 |
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(DE-627)DOAJ020422164 (DE-599)DOAJae2c4456054541f9aec0c4f9751bc98b DE-627 ger DE-627 rakwb eng Sarah J. Morgan verfasserin aut A population-level strain genotyping method to study pathogen strain dynamics in human infections 2021 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier A hallmark of chronic bacterial infections is the long-term persistence of 1 or more pathogen species at the compromised site. Repeated detection of the same bacterial species can suggest that a single strain or lineage is continually present. However, infection with multiple strains of a given species, strain acquisition and loss, and changes in strain relative abundance can occur. Detecting strain-level changes and their effects on disease is challenging because most methods require labor-intensive isolate-by-isolate analyses, and thus, only a few cells from large infecting populations can be examined. Here, we present a population-level method for enumerating and measuring the relative abundance of strains called population multi-locus sequence typing (PopMLST). The method exploits PCR amplification of strain-identifying polymorphic loci, next-generation sequencing to measure allelic variants, and informatic methods to determine whether variants arise from sequencing errors or low-abundance strains. These features enable PopMLST to simultaneously interrogate hundreds of bacterial cells that are cultured en masse from patient samples or are present in DNA directly extracted from clinical specimens without ex vivo culture. This method could be used to detect epidemic or super-infecting strains, facilitate understanding of strain dynamics during chronic infections, and enable studies that link strain changes to clinical outcomes. Infectious disease Microbiology Medicine R Samantha L. Durfey verfasserin aut Sumedha Ravishankar verfasserin aut Peter Jorth verfasserin aut Wendy Ni verfasserin aut Duncan T. Skerrett verfasserin aut Moira L. Aitken verfasserin aut Edward F. McKone verfasserin aut Stephen J. Salipante verfasserin aut Matthew C. Radey verfasserin aut Pradeep K. Singh verfasserin aut In JCI Insight American Society for Clinical investigation, 2020 6(2021), 24 (DE-627)872610594 (DE-600)2874757-4 23793708 nnns volume:6 year:2021 number:24 https://doaj.org/article/ae2c4456054541f9aec0c4f9751bc98b kostenfrei https://doi.org/10.1172/jci.insight.152472 kostenfrei https://doaj.org/toc/2379-3708 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 6 2021 24 |
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(DE-627)DOAJ020422164 (DE-599)DOAJae2c4456054541f9aec0c4f9751bc98b DE-627 ger DE-627 rakwb eng Sarah J. Morgan verfasserin aut A population-level strain genotyping method to study pathogen strain dynamics in human infections 2021 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier A hallmark of chronic bacterial infections is the long-term persistence of 1 or more pathogen species at the compromised site. Repeated detection of the same bacterial species can suggest that a single strain or lineage is continually present. However, infection with multiple strains of a given species, strain acquisition and loss, and changes in strain relative abundance can occur. Detecting strain-level changes and their effects on disease is challenging because most methods require labor-intensive isolate-by-isolate analyses, and thus, only a few cells from large infecting populations can be examined. Here, we present a population-level method for enumerating and measuring the relative abundance of strains called population multi-locus sequence typing (PopMLST). The method exploits PCR amplification of strain-identifying polymorphic loci, next-generation sequencing to measure allelic variants, and informatic methods to determine whether variants arise from sequencing errors or low-abundance strains. These features enable PopMLST to simultaneously interrogate hundreds of bacterial cells that are cultured en masse from patient samples or are present in DNA directly extracted from clinical specimens without ex vivo culture. This method could be used to detect epidemic or super-infecting strains, facilitate understanding of strain dynamics during chronic infections, and enable studies that link strain changes to clinical outcomes. Infectious disease Microbiology Medicine R Samantha L. Durfey verfasserin aut Sumedha Ravishankar verfasserin aut Peter Jorth verfasserin aut Wendy Ni verfasserin aut Duncan T. Skerrett verfasserin aut Moira L. Aitken verfasserin aut Edward F. McKone verfasserin aut Stephen J. Salipante verfasserin aut Matthew C. Radey verfasserin aut Pradeep K. Singh verfasserin aut In JCI Insight American Society for Clinical investigation, 2020 6(2021), 24 (DE-627)872610594 (DE-600)2874757-4 23793708 nnns volume:6 year:2021 number:24 https://doaj.org/article/ae2c4456054541f9aec0c4f9751bc98b kostenfrei https://doi.org/10.1172/jci.insight.152472 kostenfrei https://doaj.org/toc/2379-3708 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 6 2021 24 |
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(DE-627)DOAJ020422164 (DE-599)DOAJae2c4456054541f9aec0c4f9751bc98b DE-627 ger DE-627 rakwb eng Sarah J. Morgan verfasserin aut A population-level strain genotyping method to study pathogen strain dynamics in human infections 2021 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier A hallmark of chronic bacterial infections is the long-term persistence of 1 or more pathogen species at the compromised site. Repeated detection of the same bacterial species can suggest that a single strain or lineage is continually present. However, infection with multiple strains of a given species, strain acquisition and loss, and changes in strain relative abundance can occur. Detecting strain-level changes and their effects on disease is challenging because most methods require labor-intensive isolate-by-isolate analyses, and thus, only a few cells from large infecting populations can be examined. Here, we present a population-level method for enumerating and measuring the relative abundance of strains called population multi-locus sequence typing (PopMLST). The method exploits PCR amplification of strain-identifying polymorphic loci, next-generation sequencing to measure allelic variants, and informatic methods to determine whether variants arise from sequencing errors or low-abundance strains. These features enable PopMLST to simultaneously interrogate hundreds of bacterial cells that are cultured en masse from patient samples or are present in DNA directly extracted from clinical specimens without ex vivo culture. This method could be used to detect epidemic or super-infecting strains, facilitate understanding of strain dynamics during chronic infections, and enable studies that link strain changes to clinical outcomes. Infectious disease Microbiology Medicine R Samantha L. Durfey verfasserin aut Sumedha Ravishankar verfasserin aut Peter Jorth verfasserin aut Wendy Ni verfasserin aut Duncan T. Skerrett verfasserin aut Moira L. Aitken verfasserin aut Edward F. McKone verfasserin aut Stephen J. Salipante verfasserin aut Matthew C. Radey verfasserin aut Pradeep K. Singh verfasserin aut In JCI Insight American Society for Clinical investigation, 2020 6(2021), 24 (DE-627)872610594 (DE-600)2874757-4 23793708 nnns volume:6 year:2021 number:24 https://doaj.org/article/ae2c4456054541f9aec0c4f9751bc98b kostenfrei https://doi.org/10.1172/jci.insight.152472 kostenfrei https://doaj.org/toc/2379-3708 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 6 2021 24 |
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(DE-627)DOAJ020422164 (DE-599)DOAJae2c4456054541f9aec0c4f9751bc98b DE-627 ger DE-627 rakwb eng Sarah J. Morgan verfasserin aut A population-level strain genotyping method to study pathogen strain dynamics in human infections 2021 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier A hallmark of chronic bacterial infections is the long-term persistence of 1 or more pathogen species at the compromised site. Repeated detection of the same bacterial species can suggest that a single strain or lineage is continually present. However, infection with multiple strains of a given species, strain acquisition and loss, and changes in strain relative abundance can occur. Detecting strain-level changes and their effects on disease is challenging because most methods require labor-intensive isolate-by-isolate analyses, and thus, only a few cells from large infecting populations can be examined. Here, we present a population-level method for enumerating and measuring the relative abundance of strains called population multi-locus sequence typing (PopMLST). The method exploits PCR amplification of strain-identifying polymorphic loci, next-generation sequencing to measure allelic variants, and informatic methods to determine whether variants arise from sequencing errors or low-abundance strains. These features enable PopMLST to simultaneously interrogate hundreds of bacterial cells that are cultured en masse from patient samples or are present in DNA directly extracted from clinical specimens without ex vivo culture. This method could be used to detect epidemic or super-infecting strains, facilitate understanding of strain dynamics during chronic infections, and enable studies that link strain changes to clinical outcomes. Infectious disease Microbiology Medicine R Samantha L. Durfey verfasserin aut Sumedha Ravishankar verfasserin aut Peter Jorth verfasserin aut Wendy Ni verfasserin aut Duncan T. Skerrett verfasserin aut Moira L. Aitken verfasserin aut Edward F. McKone verfasserin aut Stephen J. Salipante verfasserin aut Matthew C. Radey verfasserin aut Pradeep K. Singh verfasserin aut In JCI Insight American Society for Clinical investigation, 2020 6(2021), 24 (DE-627)872610594 (DE-600)2874757-4 23793708 nnns volume:6 year:2021 number:24 https://doaj.org/article/ae2c4456054541f9aec0c4f9751bc98b kostenfrei https://doi.org/10.1172/jci.insight.152472 kostenfrei https://doaj.org/toc/2379-3708 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 6 2021 24 |
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A population-level strain genotyping method to study pathogen strain dynamics in human infections |
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A hallmark of chronic bacterial infections is the long-term persistence of 1 or more pathogen species at the compromised site. Repeated detection of the same bacterial species can suggest that a single strain or lineage is continually present. However, infection with multiple strains of a given species, strain acquisition and loss, and changes in strain relative abundance can occur. Detecting strain-level changes and their effects on disease is challenging because most methods require labor-intensive isolate-by-isolate analyses, and thus, only a few cells from large infecting populations can be examined. Here, we present a population-level method for enumerating and measuring the relative abundance of strains called population multi-locus sequence typing (PopMLST). The method exploits PCR amplification of strain-identifying polymorphic loci, next-generation sequencing to measure allelic variants, and informatic methods to determine whether variants arise from sequencing errors or low-abundance strains. These features enable PopMLST to simultaneously interrogate hundreds of bacterial cells that are cultured en masse from patient samples or are present in DNA directly extracted from clinical specimens without ex vivo culture. This method could be used to detect epidemic or super-infecting strains, facilitate understanding of strain dynamics during chronic infections, and enable studies that link strain changes to clinical outcomes. |
abstractGer |
A hallmark of chronic bacterial infections is the long-term persistence of 1 or more pathogen species at the compromised site. Repeated detection of the same bacterial species can suggest that a single strain or lineage is continually present. However, infection with multiple strains of a given species, strain acquisition and loss, and changes in strain relative abundance can occur. Detecting strain-level changes and their effects on disease is challenging because most methods require labor-intensive isolate-by-isolate analyses, and thus, only a few cells from large infecting populations can be examined. Here, we present a population-level method for enumerating and measuring the relative abundance of strains called population multi-locus sequence typing (PopMLST). The method exploits PCR amplification of strain-identifying polymorphic loci, next-generation sequencing to measure allelic variants, and informatic methods to determine whether variants arise from sequencing errors or low-abundance strains. These features enable PopMLST to simultaneously interrogate hundreds of bacterial cells that are cultured en masse from patient samples or are present in DNA directly extracted from clinical specimens without ex vivo culture. This method could be used to detect epidemic or super-infecting strains, facilitate understanding of strain dynamics during chronic infections, and enable studies that link strain changes to clinical outcomes. |
abstract_unstemmed |
A hallmark of chronic bacterial infections is the long-term persistence of 1 or more pathogen species at the compromised site. Repeated detection of the same bacterial species can suggest that a single strain or lineage is continually present. However, infection with multiple strains of a given species, strain acquisition and loss, and changes in strain relative abundance can occur. Detecting strain-level changes and their effects on disease is challenging because most methods require labor-intensive isolate-by-isolate analyses, and thus, only a few cells from large infecting populations can be examined. Here, we present a population-level method for enumerating and measuring the relative abundance of strains called population multi-locus sequence typing (PopMLST). The method exploits PCR amplification of strain-identifying polymorphic loci, next-generation sequencing to measure allelic variants, and informatic methods to determine whether variants arise from sequencing errors or low-abundance strains. These features enable PopMLST to simultaneously interrogate hundreds of bacterial cells that are cultured en masse from patient samples or are present in DNA directly extracted from clinical specimens without ex vivo culture. This method could be used to detect epidemic or super-infecting strains, facilitate understanding of strain dynamics during chronic infections, and enable studies that link strain changes to clinical outcomes. |
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A population-level strain genotyping method to study pathogen strain dynamics in human infections |
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