Comparison of<i>dkgB</i>-linked intergenic sequence ribotyping to DNA microarray hybridization for assigning serotype to<i>Salmonella enterica</i>

Guard, Jean; Sanchez-Ingunza, Roxana; Morales, Cesar A.; Stewart, Tod; Liljebjelke, Karen; Kessel, JoAnn Van; Ingram, Kim; Jones, D. R.; Jackson, Charlene R.; Fedorka–Cray, Paula J.; Frye, Jonathan G.; Gast, Richard K.; Hinton, Arthur

doi:10.1111/1574-6968.12010

Cited by 32 publications

(29 citation statements)

References 11 publications

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“…As the cost of 7-gene MLST has been estimated at Ͻ$35 per isolate (24) when typing ϳ200 isolates per week, the molecular serotyping procedure described here, which involves sequencing of 2 genes plus up to three multiplex PCRs, should only cost about $15 to $20 per isolate. This is less expensive than traditional serotyping (estimated at $35 to $185 per isolate) (20).…”

Section: Discussionmentioning

confidence: 99%

“…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%

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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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Section: Discussionmentioning

confidence: 99%

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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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“…The European Centre for Disease Prevention and Control (ECDC) recently reported a median cost of €29 ($43 CAD) to serotype a common Salmonella serovar, including culturing and higher costs associated with more difficult cases (21). There are reports of traditional serotyping costs as high as $185 USD (22), but it is difficult to assess this wide range in costs as it depends on methods used, whether antisera is purchased or produced in-house, and whether the estimate includes labor. The cost of WGS is up to three times more expensive than molecular serotyping methods at $132 CAD (A. Reimer, personal communication, April 2016).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Molecular Methods for Identification of Salmonella Serovars

et al. 2016

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bClassification by serotyping is the essential first step in the characterization of Salmonella isolates and is important for surveillance, source tracking, and outbreak detection. To improve detection and reduce the burden of salmonellosis, several rapid and high-throughput molecular Salmonella serotyping methods have been developed.The aim of this study was to compare three commercial kits, Salm SeroGen (Salm Sero-Genotyping AS-1 kit), Check&Trace (Check-Points), and xMAP (xMAP Salmonella serotyping assay), to the Salmonella genoserotyping array (SGSA) developed by our laboratory. They were assessed using a panel of 321 isolates that represent commonly reported serovars from human and nonhuman sources globally. The four methods correctly identified 73.8% to 94.7% of the isolates tested. The methods correctly identified 85% and 98% of the clinically important Salmonella serovars Enteritidis and Typhimurium, respectively. The methods correctly identified 75% to 100% of the nontyphoidal, broad host range Salmonella serovars, including Heidelberg, Hadar, Infantis, Kentucky, Montevideo, Newport, and Virchow. The sensitivity and specificity of Salmonella serovars Typhimurium and Enteritidis ranged from 85% to 100% and 99% to 100%, respectively.It is anticipated that whole-genome sequencing will replace serotyping in public health laboratories in the future. However, at present, it is approximately three times more expensive than molecular methods. Until consistent standards and methodologies are deployed for whole-genome sequencing, data analysis and interlaboratory comparability remain a challenge. The use of molecular serotyping will provide a valuable high-throughput alternative to traditional serotyping. This comprehensive analysis provides a detailed comparison of commercial kits available for the molecular serotyping of Salmonella.

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“…The digested genomic DNA of target bacteria is separated on an agarose gel and then hybridized with complementary sequences for identifying the banding patern. A database of ingerprint species, serovar, and strain identiications is used for comparison [21][22][23]. The ingerprinting methods include pulsed-ield gel electrophoresis (PFGE), ribotyping, and intergenic sequence (IGS) ribotyping.…”

Section: Culture-dependent Methodsmentioning

confidence: 99%

Current and Emerging Innovations for Detection of Food-Borne Salmonella

Wu¹,

Zeng²

2017

Current Topics in Salmonella and Salmonellosis

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Salmonella is one of the leading causes of food-borne illnesses worldwide, and one of the main contributors to salmonellosis is the consumption of contaminated egg, poultry, pork, beef, and milk products. Since deleterious efects of Salmonella on public health and the economy continue to occur, improving safety of food products by early detection of food-borne pathogens would be considered an important component for limiting exposure to Salmonella contamination. Therefore, there is an ongoing need to develop more advanced detection methods that can identify Salmonella accurately and rapidly in foods before they reach consumers. In the past three decades, there have been increasing eforts toward developing and improving rapid pathogen detection and characterization methodologies for application to food products. In this chapter, we discuss molecular methods for detection, identiication, and genetic characterization of Salmonella in food. In addition, the advantages and disadvantages of the established and emerging rapid detection methods are addressed here. The methods with potential application to the industry are highlighted in this chapter.

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Comparison ofdkgB-linked intergenic sequence ribotyping to DNA microarray hybridization for assigning serotype toSalmonella enterica

Cited by 32 publications

References 11 publications

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

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

Evaluation of Molecular Methods for Identification of Salmonella Serovars

Current and Emerging Innovations for Detection of Food-Borne Salmonella

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