Determination of Escherichia coli O types by allele-specific polymerase chain reaction: application to the O types involved in human septicemia

Olivier Clermont; James R. Johnson; Megan Menard; Erick Denamur

doi:10.1016/j.diagmicrobio.2006.08.007

Determination of Escherichia coli O types by allele-specific polymerase chain reaction: application to the O types involved in human septicemia

Olivier Clermont, James R. Johnson, Megan Menard, Erick Denamur

Medicine - Veteran's Administration Medical Center

Research output: Contribution to journal › Article › peer-review

111 Scopus citations

Abstract

Escherichia coli can be serotyped by determination of somatic (O), capsular (K), and flagellar (H) antigens, and clear associations exist between specific O antigens and pathogenic behavior. However, E. coli is very challenging to O type with traditional serologic methods, making new methods for E. coli somatic antigen detection highly desirable. Here, we describe a simple alternative molecular method for determination of the E. coli O type based on allele-specific polymerase chain reaction amplification of the 5′ portion of the rfb locus. We present our application of this new method to the detection of the 12 principal O types (O1, O2, O4, O6, O7, O12, O15, O16, O18, O25, O75, and O157) found among bloodstream isolates of E. coli. This method allowed us to determine the O types of 153 strains previously typed using reference methods with an accuracy exceeding 90%. Moreover, some rough or nonagglutinating strains can be successfully typed.

Original language	English (US)
Pages (from-to)	129-136
Number of pages	8
Journal	Diagnostic Microbiology and Infectious Disease
Volume	57
Issue number	2
DOIs	https://doi.org/10.1016/j.diagmicrobio.2006.08.007
State	Published - Feb 2007

Bibliographical note

Funding Information:
The authors are grateful to Stéphanie Gouriou, Catherine Branger, and Bertrand Picard for providing some of the strains used in this study and to Stéphane Bonacorsi and Edouard Bingen for discussions in the early conception of the assay. This work was supported in part by the “Fondation pour la Recherche Médicale” (E.D.) and Office of Research and Development, Medical Research Service, Department of Veterans Affairs, Minneapolis, MN (J.R.J.).

Keywords

Allele specific PCR
Bloodstream isolates
Escherichia coli
Serotype O

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title = "Determination of Escherichia coli O types by allele-specific polymerase chain reaction: application to the O types involved in human septicemia",

abstract = "Escherichia coli can be serotyped by determination of somatic (O), capsular (K), and flagellar (H) antigens, and clear associations exist between specific O antigens and pathogenic behavior. However, E. coli is very challenging to O type with traditional serologic methods, making new methods for E. coli somatic antigen detection highly desirable. Here, we describe a simple alternative molecular method for determination of the E. coli O type based on allele-specific polymerase chain reaction amplification of the 5′ portion of the rfb locus. We present our application of this new method to the detection of the 12 principal O types (O1, O2, O4, O6, O7, O12, O15, O16, O18, O25, O75, and O157) found among bloodstream isolates of E. coli. This method allowed us to determine the O types of 153 strains previously typed using reference methods with an accuracy exceeding 90%. Moreover, some rough or nonagglutinating strains can be successfully typed.",

keywords = "Allele specific PCR, Bloodstream isolates, Escherichia coli, Serotype O",

author = "Olivier Clermont and Johnson, {James R.} and Megan Menard and Erick Denamur",

note = "Funding Information: The authors are grateful to St{\'e}phanie Gouriou, Catherine Branger, and Bertrand Picard for providing some of the strains used in this study and to St{\'e}phane Bonacorsi and Edouard Bingen for discussions in the early conception of the assay. This work was supported in part by the “Fondation pour la Recherche M{\'e}dicale” (E.D.) and Office of Research and Development, Medical Research Service, Department of Veterans Affairs, Minneapolis, MN (J.R.J.).",

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AU - Denamur, Erick

N1 - Funding Information: The authors are grateful to Stéphanie Gouriou, Catherine Branger, and Bertrand Picard for providing some of the strains used in this study and to Stéphane Bonacorsi and Edouard Bingen for discussions in the early conception of the assay. This work was supported in part by the “Fondation pour la Recherche Médicale” (E.D.) and Office of Research and Development, Medical Research Service, Department of Veterans Affairs, Minneapolis, MN (J.R.J.).

PY - 2007/2

Y1 - 2007/2

N2 - Escherichia coli can be serotyped by determination of somatic (O), capsular (K), and flagellar (H) antigens, and clear associations exist between specific O antigens and pathogenic behavior. However, E. coli is very challenging to O type with traditional serologic methods, making new methods for E. coli somatic antigen detection highly desirable. Here, we describe a simple alternative molecular method for determination of the E. coli O type based on allele-specific polymerase chain reaction amplification of the 5′ portion of the rfb locus. We present our application of this new method to the detection of the 12 principal O types (O1, O2, O4, O6, O7, O12, O15, O16, O18, O25, O75, and O157) found among bloodstream isolates of E. coli. This method allowed us to determine the O types of 153 strains previously typed using reference methods with an accuracy exceeding 90%. Moreover, some rough or nonagglutinating strains can be successfully typed.

AB - Escherichia coli can be serotyped by determination of somatic (O), capsular (K), and flagellar (H) antigens, and clear associations exist between specific O antigens and pathogenic behavior. However, E. coli is very challenging to O type with traditional serologic methods, making new methods for E. coli somatic antigen detection highly desirable. Here, we describe a simple alternative molecular method for determination of the E. coli O type based on allele-specific polymerase chain reaction amplification of the 5′ portion of the rfb locus. We present our application of this new method to the detection of the 12 principal O types (O1, O2, O4, O6, O7, O12, O15, O16, O18, O25, O75, and O157) found among bloodstream isolates of E. coli. This method allowed us to determine the O types of 153 strains previously typed using reference methods with an accuracy exceeding 90%. Moreover, some rough or nonagglutinating strains can be successfully typed.

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KW - Serotype O

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Determination of Escherichia coli O types by allele-specific polymerase chain reaction: application to the O types involved in human septicemia

Abstract

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