Principles and Applications of Methods for DNA-Based Typing of Microbial Organisms

Olive, D; Bean, Pamela

doi:10.1128/jcm.37.6.1661-1669.1999

Cited by 750 publications

(320 citation statements)

References 96 publications

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“…These results could be expected since these methods, despite being widely used for common purposes, rely on the analysis of distinct targets, i.e. PFGE and RAPD-PCR are current methods based on variations that evolve very rapidly and MLEE makes use of variations that are slowly accumulated [16,24,38]. Despite the high polymorphism detected by PFGE and RAPD, genetic exchanges of virulence determinants in the studied population seem to occur among phylogenetically related E. coli strains as revealed by the MLEE clustering linkage.…”

Section: Discussionmentioning

confidence: 99%

Genotypic and phenotypic characterization of enterotoxigenicEscherichia coli(ETEC) strains isolated in Rio de Janeiro city, Brazil

Régua-Mangia

Guth

Andrade

et al. 2004

FEMS Immunology &Amp; Medical Microbiology

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Enterotoxigenic Escherichia coli (ETEC) strains have been implicated as important etiological agents of diarrheal disease, especially in developing countries. This group of microorganisms has been associated with a diverse range of genotypic and phenotypic markers. In the present study, 21 ETEC isolates previously defined according to the toxigenic genotypes, were characterized on the basis of O:H typing, cell adherence patterns, and colonization factors (CFs) antigens. Genetic diversity was investigated by random amplification polymorphic DNA (RAPD-PCR), pulsed-field gel electrophoresis (PFGE) and multilocus enzyme electrophoresis (MLEE). LT-I probe-positive isolates belonged to serotypes ONT:HNT, O7:H24, O48:H21, O88:H25, O148:H28, O159:H17 and O159:H21. ST-h probe-positive isolates belonged to serotypes O159:H17, O148:H28 and O6:H-. Serotypes O148:H28, O159:H17 and O6:H- were associated with the CS6, CFA/I and CS1 CS3 antigens, respectively. Most ETEC strains exhibited a diffuse pattern of adherence to cultured epithelial cells. In general, phenotypic and genotypic characteristics correlated well. RAPD-PCR, PFGE and MLEE showed reproducibility and good discriminatory potential. The application of molecular typing systems allowed the detection of significant diversity among the isolates, indicating a non-clonal origin and revealing intra-serotype variation overlooked by classical epidemiological approaches. The phenotypic and genotypic diversity observed lead us to recommend the use of different typing systems in order to elucidate the epidemiology of ETEC infection.

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Section: Discussionmentioning

confidence: 99%

Genotypic and phenotypic characterization of enterotoxigenicEscherichia coli(ETEC) strains isolated in Rio de Janeiro city, Brazil

Régua-Mangia

Guth

Andrade

et al. 2004

FEMS Immunology &Amp; Medical Microbiology

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“…Based on these findings, it can be concluded that the use of ERIC-PCR and REP-PCR are beneficial, and these approaches have shown to be useful for assessing the genetic diversity of isolates from different geographic locations (Mehta et al, 2002). Moreover, the consumables and labor costs for ERIC-PCR and REP-PCR analyses are significantly lower than those for other genotyping methods (Olive and Bean, 1999).…”

Section: Eric-pcrmentioning

confidence: 94%

Source identification of airborne Escherichia coli of swine house surroundings using ERIC-PCR and REP-PCR

Duan

Chai

Liu

et al. 2009

Environmental Research

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Evidence is mounting that microorganisms originating from livestock impact the air quality of the animal houses themselves and the public in the surrounding neighborhoods. The aim of this study was to develop efficient bacterial source tracking capabilities to identify sources of Escherichia coli aerosol pollution caused by pigs. Airborne E. coli were isolated from indoor air, upwind air (10 and 50 m away) and downwind air samples (10, 50, 100, 200 and 400 m away) for five swine houses using six-stage Andersen microbial samplers and Reuter-Centrifugal samplers (RCS). E. coli strains from pig fecal samples were also collected simultaneously. The enterobacterial repetitive intergenic consensus polymerize chain reaction (ERIC-PCR) and the repetitive extragenic palindromic (REP-PCR) approaches were used to study the genetic variability and to determine the strain relationships among E. coli isolated from different sites in each swine house. Results showed that 35.1% (20/57) of the bacterial DNA fingerprints from the fecal isolates matched with the corresponding strains isolated from indoor and downwind air samples (similarity > or = 90%). E. coli strains from the indoor and downwind air samples were closely related to the E. coli strains isolated from feces, while those isolated from upwind air samples (swine house C) had low similarity (61-69%). Our results suggest that some strains isolated from downwind and indoor air originated in the swine feces. Effective hygienic measures should be taken in animal farms to prevent or minimize the downwind spread of microorganism aerosol.

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“…Generally, higher discriminatory power, speed, standardization, and ease of use are associated with a higher cost. All of these factors, the availability of equipment and trained personnel, and most important, the epidemiologic question determine the suitability of a typing technique for any given situation [29,30].…”

Section: Molecular Methodsmentioning

confidence: 99%

Use of Molecular Epidemiology in Veterinary Practice

Zadoks

Schukken

2006

Veterinary Clinics of North America: Food Animal Practice

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Molecular epidemiology is a relatively new branch of epidemiology that uses molecular biology methods to study health and disease in populations. This article gives an introduction to molecular epidemiologic terminology and methodology and its usefulness in large animal medicine and veterinary public health. Applications in source tracing and vaccine studies and insights into transmission dynamics, host specificity, and niche adaptation of infectious organisms are presented. Examples are drawn from a variety of diseases, organisms, and host species and range from the global level to the individual-animal level.

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Principles and Applications of Methods for DNA-Based Typing of Microbial Organisms

Cited by 750 publications

References 96 publications

Genotypic and phenotypic characterization of enterotoxigenicEscherichia coli(ETEC) strains isolated in Rio de Janeiro city, Brazil

Genotypic and phenotypic characterization of enterotoxigenicEscherichia coli(ETEC) strains isolated in Rio de Janeiro city, Brazil

Source identification of airborne Escherichia coli of swine house surroundings using ERIC-PCR and REP-PCR

Use of Molecular Epidemiology in Veterinary Practice

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