Rapid Genoserotyping Tool for Classification of Salmonella Serovars

Franklin, Kristyn; Lingohr, Erika J.; Yoshida, Catherine; Anjum, Muna F.; Bodrossy, Levente; Clark, Clifford G.; Kropinski, Andrew M.; Karmali, Mohamed A.

doi:10.1128/jcm.02347-10

Cited by 51 publications

(46 citation statements)

References 59 publications

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“…In most Salmonella strains, flagellin expression is coordinately expressed via a phase-variation mechanism (12). A number of studies have utilized the variabilities of the rfb region, fliC, and fljB to identify serovars, typically using probe-based assays or PCR strategies (13)(14)(15). While these approaches have been reported to show good concordance with traditional serotyping, the limitations of these methods include problems with the characterization of new or unusual serovars or allelic variants that do not react with existing primers or probes (13)(14)(15).…”

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

et al. 2013

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“…DNA microarray is a high-throughput technology for simultaneous determination of the presence or absence of a large number of genes, including housekeeping genes and virulence genes [63][64][65][66][67][68]. DNA microarray has been employed to detect and identify various bacterial pathogens in food in one experiment.…”

Section: Dna Microarraymentioning

confidence: 99%

Recent and the Latest Developments in Rapid and Efficient Detection of Salmonella in Food and Water

Hu¹,

Li²

2017

Adv Tech Biol Med

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Salmonella is a leading cause of foodborne diseases. Gastroenteritis caused by nontyphoid Salmonella is still a major infectious disease in the world. About 95% of Salmonella infections are caused by ingestion of contaminated food and water. Rapid, sensitive, and efficient detection and identification of Salmonella from foods and water are critical for minimizing the spread of outbreaks caused by this pathogen. These methods can be applied to track the food source of contamination or by early diagnosis of the infections in a clinical setting. Culture-based methods for detection of Salmonella are laborious and time-consuming, typically taking 5-7 days to obtain a pure culture and serovar identification. In the past few decades, molecular-based technologies have greatly shortened the time for detection and identification of bacterial pathogens from food and water, and drastically increased the specificity and sensitivity of the assays. In this review, we report an update on the development of rapid and efficient methods for the detection and identification of Salmonella in food and water, focusing on the approaches for concentration of Salmonella cells and viable cell detection, as well as the advantages and drawbacks of these methods.

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“…Although serotyping and PFGE have held their place in the reference laboratories for decades, there are drawbacks to each typing method. Both methods are time-consuming and require expensive reagents, equipment, and highly trained staff (6,7). However, due to the wealth of information collected from traditional serotyping and PFGE, it is important that new high-throughput molecular methods produce data comparable to historical data.…”

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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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Rapid Genoserotyping Tool for Classification of Salmonella Serovars

Cited by 51 publications

References 59 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

Recent and the Latest Developments in Rapid and Efficient Detection of Salmonella in Food and Water

Evaluation of Molecular Methods for Identification of Salmonella Serovars

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