Pulsed-Field Gel Electrophoresis Subtyping Database for Foodborne<i>Salmonella enterica</i>Serotype Discrimination

Kérouanton, Annaëlle; Marault, Muriel; Lailler, Renaud; Weill, François‐Xavier; Feurer, Carole; Espié, Emmanuelle; Brisabois, Anne

doi:10.1089/fpd.2007.0090

Cited by 59 publications

(59 citation statements)

References 44 publications

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“…PFGE produces comparable data of genotypic characteristics of Salmonella strains and it is accepted as the gold standard among molecular methods [12,13]. In addition to this, it has been used in typing Salmonella in human patients, animal sources and foods because of its discriminatory power and high reproducibility [14][15][16]. Numerous reports have been documented that using the highly discriminatory technique of PFGE was successful to track the source of Salmonella infections in different serovars [17][18][19][20][21].…”

Section: Introductionmentioning

confidence: 99%

“…Numerous reports have been documented that using the highly discriminatory technique of PFGE was successful to track the source of Salmonella infections in different serovars [17][18][19][20][21]. However, PFGE does not display equal sensitivity in different serovars [14,21] and it has several limitations including changes in DNA concentration, percent of agarose in the gel, applied voltage and gel temperature. Beside these, it requires at least 3-4 days labour intensive to complete the test and the presence of expensive specialized equipment and high quality chemicals [7,22,23].…”

Section: Introductionmentioning

confidence: 99%

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Genetic diversity of food originated Salmonella isolates

Karatuğ

Yüksel

Akçelik

et al. 2018

Biotechnology & Biotechnological Equipment

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Currently, Salmonella enterica is the most common bacterial foodborne pathogen, causing serious extraintestinal disease. Typing methods play an important role on pathogens' source tracking, knowing the source(s) of bacteria in pharmaceutical sciences, preventing and controlling the diarrhea and food-poisoning outbreaks. The purpose of this study is to use different moleculer typing methods to determine the genetic variability of 38 foodborne Salmonella isolates that were previously identified by biochemical tests. The methods were evaluated by four molecular techniques including 16S rRNA sequencing, PFGE, PCR-RFLP and invA-spvC PCR. 16S rRNA sequencing results showed that four of the 38 isolates were Escherichia coli, Proteus mirabilis and Citrobacter murliniae, and the others were Salmonella enterica. Thirty-eight strains were subtyped by XbaI-PFGE into 20 profiles with different clusters, while they were subtyped by 16S rRNA-RFLP into 9 profiles with a single cluster. Out of two Salmonella isolates, the invasion gene (invA) was detected in all other Salmonella isolates (94%) and the virulence gene (spvC) was detected in 11% of Salmonella isolates. Our results suggested that the PFGE subtyping is the prominent method for the evaluation and benchmarking of molecular subtyping.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Genetic diversity of food originated Salmonella isolates

Karatuğ

Yüksel

Akçelik

et al. 2018

Biotechnology & Biotechnological Equipment

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“…In addition to PFGE (16,17), different ribotyping approaches (18)(19)(20), repetitive extragenic palindromic sequencebased PCR (rep-PCR) (21,22), multilocus sequence typing (MLST) (23,24), and molecular typing based on genomic markers (25)(26)(27)(28) have been investigated for their abilities to replace or complement traditional serotyping. While many of these methods have been able to reliably predict a limited set of serovars, they still lack widespread adoption, likely due to the requirements for specialized equipment, as well as a lack of proven reliability for predicting Salmonella serovars.…”

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confidence: 99%

Comparison of Typing Methods with a New Procedure Based on Sequence Characterization for Salmonella Serovar Prediction

et al. 2013

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bAs the development of molecular serotyping approaches is critical for Salmonella spp., which include >2,600 serovars, we performed an initial evaluation of the ability to identify Salmonella serovars using (i) different molecular subtyping methods and (ii) a newly implemented combined PCR-and sequencing-based approach that directly targets O-and H-antigen-encoding genes. Initial testing was performed using 46 isolates that represent the top 40 Salmonella serovars isolated from human and nonhuman sources, as reported by the U.S. Centers for Disease Control and Prevention and the World Health Organization. Multilocus sequence typing (MLST) was able to accurately predict the serovars for 42/46 isolates and showed the best ability to predict serovars among the subtyping methods tested. Pulsed-field gel electrophoresis (PFGE), ribotyping, and repetitive extragenic palindromic sequence-based PCR (rep-PCR) were able to accurately predict the serovars for 35/46, 34/46, and 30/46 isolates, respectively. Among the methods, S. enterica subsp. enterica serovars 4,5,12:i:؊, Typhimurium, and Typhimurium var. 5؊ were frequently not classified correctly, which is consistent with their close phylogenetic relationship. To develop a PCR-and sequence-based serotyping approach, we integrated available data sources to implement a combination PCR-based O-antigen screening and sequencing of internal fliC and fljB fragments. This approach correctly identified the serovars for 42/46 isolates in the initial set representing the most common Salmonella serovars, as well as for 54/63 isolates representing less common Salmonella serovars. Our study not only indicates that different molecular approaches show the potential to allow for rapid serovar classification of Salmonella isolates, but it also provides data that can help with the selection of molecular serotyping methods to be used by different laboratories.

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“…A number of recent strategies have employed PCR-based approaches to determine different O and H antigens as a means to replace serologic identification of these antigens (14,20,25,32). Others have proposed alternative strategies examining genetic differences as a means of identifying serovars, including ribotyping (17), pulsed-field gel electrophoresis (PFGE) (28), multiplex PCR (3,4,29), IS200 analysis (18,46), random amplification of DNA polymorphisms (45), and DNA microarray analysis (38,42).…”

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confidence: 99%

High-Throughput Molecular Determination of Salmonella enterica Serovars by Use of Multiplex PCR and Capillary Electrophoresis Analysis

Leader

Frye²,

et al. 2009

J Clin Microbiol

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Salmonella enterica is a leading cause of food-borne illness worldwide and is also a major cause of morbidity and mortality in domestic and wild animals. In the current study, a high-throughput molecular assay was developed to determine the most common clinical and nonhuman serovars of S. enterica in the United States. Sixteen genomic targets were identified based on their differential distribution among common serovars. Primers were designed to amplify regions of each of these targets in a single multiplex PCR while incorporating a 6-carboxyfluorescein-labeled universal primer to fluorescently label all amplicons. The fluorescently labeled PCR products were separated using capillary electrophoresis, and a Salmonella multiplex assay for rapid typing (SMART) code was generated for each isolate, based upon the presence or absence of PCR products generated from each target gene. Seven hundred fifty-one blind clinical isolates of Salmonella from Washington State, collected in 2007 and previously serotyped via antisera, were screened with the assay. A total of 89.6% of the isolates were correctly identified based on comparison to a panel of representative SMART codes previously determined for the top 50 most common serovars in the United States. Of the remaining isolates, 6.2% represented isolates that produced a new SMART code for a previously determined serotype, while the final 8.8% were from serotypes not screened in the original panel used to score isolates in the blinded study. This high-throughput multiplex PCR assay allowed simple and accurate typing of the most prevalent clinical serovars of Salmonella enterica at a level comparable to that of conventional serotyping, but at a fraction of both the cost and time required per test.

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Pulsed-Field Gel Electrophoresis Subtyping Database for FoodborneSalmonella entericaSerotype Discrimination

Cited by 59 publications

References 44 publications

Genetic diversity of food originated Salmonella isolates

Genetic diversity of food originated Salmonella isolates

Comparison of Typing Methods with a New Procedure Based on Sequence Characterization for Salmonella Serovar Prediction

High-Throughput Molecular Determination of Salmonella enterica Serovars by Use of Multiplex PCR and Capillary Electrophoresis Analysis

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