Multiple-locus variable-number tandem-repeats analysis of Salmonella enterica subsp. enterica serovar Typhimurium using PCR multiplexing and multicolor capillary electrophoresis

Lindstedt, Bjørn-Arne; Vardund, Traute; Aas, Lena; Kapperud, Georg

doi:10.1016/j.mimet.2004.06.014

Cited by 252 publications

(293 citation statements)

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“…Thus, the MNR-MLST and L-SSR typing (also termed MLVA) methods should be more suitable for evolutionary studies by their nature, since they parametrically detect multiple sequence variation that accumulates slowly compared to the highly discriminative PFGE method (51). Thus, multilocus SSR typing is a good tool for epidemiological and phylog-eny studies of V. cholerae, as demonstrated with other bacterial species (22,39,41,48,78). …”

Section: Discussionmentioning

confidence: 99%

“…Larger SSRs (L-SSR) consisting of two to 9-bp motifs were successfully used to analyze variation among strains by size determination of the PCR amplicon (39,81). Thus, in the last few years multiple-locus SSR, also termed multiple-locus VNTR analysis (MLVA), has been increasingly recognized as the marker of choice for genotyping a number of pathogens such as Bacillus anthracis (39), Borrelia species (22), Salmonella enterica (48), and Enterococcus faecium (78).…”

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Vibrio cholerae Strain Typing and Phylogeny Study Based on Simple Sequence Repeats

et al. 2007

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Vibrio cholerae is the etiological agent of cholera. Its natural reservoir is the aquatic environment. To date, practical typing of V. cholerae is mainly serological and requires about 200 antisera. Simple sequence repeats (SSR), also termed VNTR (for variable number of tandem repeats), provide a source of high genomic polymorphism used in bacterial typing. Here we describe an SSR-based typing method that combines the variation in highly mutable SSR loci, with that of shorter, relatively more stable mononucleotide repeat (MNR) loci, for accurate and rapid typing of V. cholerae. Vibrio cholerae, a gram-negative bacterium, is the causal agent of the severe diarrheal disease cholera. Its natural reservoir is the aquatic environment (49, 84). Cholera pandemics are caused by specific serogroups of V. cholerae that are pathogenic only to humans (35,84). Since 1817, V. cholerae has caused a number of pandemics. The first seven were presumed to be caused by O1 serogroup of V. cholerae (68). In October 1992, a new serogroup, defined O139, caused a severe outbreak of cholera in southeast India. Within 10 months, the O139 serogroup was disseminated all over the Indian subcontinent and soon thereafter spread to 11 neighboring countries, temporarily displacing the O1 serogroups (64). Since then both serogroups coexist and are responsible for large outbreaks. Genomic studies indicated that O139 epidemic strain arose by horizontal acquisition of unique DNA (15, 47).Traditionally, V. cholerae classification is serological and requires about 200 antisera based on the somatic O antigen (72). Isolates of V. cholerae are divided into three major subgroups: O1, O139, and non-O1/non-O139, of which only the O1 and O139 serogroups are associated with cholera pandemics and epidemics. Non-O1, non-O139 serogroups are recognized as causative agents of sporadic and localized outbreaks (73). Pathogenic V. cholerae isolates carry virulence genes, such as the toxins genes ctxAB (25,65,68). The environmental V. cholerae strains from non-O1, non-O139 serogroups are a possible natural reservoir of potentially new emerging epidemic strains (67,73). This assumption is supported by the finding that some of these environmental strains harbor virulence genes (23) and thus are likely to evolve into novel pathogenic strains by horizontal gene transfer (24,25). The emergence of new pathogenic V. cholerae strains requires not only an efficient, rapid, and accurate identification tool but also a means for determining genetic relationships among environmental and clinical isolates.Genome-based bacterial identification and typing is essential for several disciplines, including taxonomy, epidemiology, determining phylogenetic relationships, and the study of evolutionary mechanisms. It allows distinguishing among strains within a species to monitor epidemics and routes of contamination. Recent advances in biotechnology have resulted in the development of numerous methods for microorganism typing that differ in their sensitivity, rapidity, complexity, discrimin...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Vibrio cholerae Strain Typing and Phylogeny Study Based on Simple Sequence Repeats

et al. 2007

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“…MLVA typing was performed as described previously by Lindstedt et al (2004), with some modifications. The modifications were a changed dye set from DS-34 to DS-33 for primer labelling.…”

Section: Methodsmentioning

confidence: 99%

Genotypic diversity, pathogenic potential and the resistance profile of Salmonella Typhimurium strains isolated from humans and food from 1983 to 2013 in Brazil

Almeida

Medeiros

Rodrigues

et al. 2015

Journal of Medical Microbiology

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Salmonella enterica subsp. enterica serovar Typhimurium is one of the leading serovars that causes salmonellosis worldwide. However, few studies have molecularly characterized S. Typhimurium strains in Brazil. In this study, we genotyped 92 S. Typhimurium strains isolated from humans (43) and food (49) between 1983 and 2013 in Brazil using PFGE, multiple-locus variable number of tandem repeats analysis (MLVA) and enterobacterial repetitive intergenic consensus PCR (ERIC-PCR). Moreover, we assessed the frequency of 12 virulence markers by PCR and the resistance profile against 12 antimicrobials. More than 85.8 % of the strains studied carried 11 of the virulence markers or more. Thirty-three strains (25 %) were multidrug resistant (MDR). The 92 S. Typhimurium studied were grouped by PFGE as PFGE-A, PFGE-B1 and PFGE-B2; by MLVA as MLVA-A, MLVA-B1 and MLVA-B2; and, finally, by ERIC-PCR as ERIC-A and ERIC-B. The strains isolated from humans before the mid-1990s were allocated to all clusters. The strains isolated from humans after the mid-1990s were distributed in the PFGE-B1, MLVA-B1, MLVA-B2 and ERIC-A clusters. The strains isolated from food were distributed in all clusters, except in PFGE-B2. All typing results suggested that the S. Typhimurium strains of human clinical origin isolated before the mid-1990s were genetically more diverse, which might indicate the selection of a more adapted S. Typhimurium subtype after Salmonella Enteritidis became the most prevalent serovar in Brazil. Regarding strains isolated from food, the results suggest the current circulation of more than one subtype. Furthermore, the high frequency of virulence genes and the presence of MDR strains reinforces their potential hazard for humans and the risk of their presence in foods in Brazil.

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“…Using labeled primers with different dyes, it is possible to perform multiplex PCR followed by CGE-LIF to provide microbiological identification with less uncertainty than with AGE analysis. This methodology has proven to be a powerful tool for differentiating serovars of S. enterica and Shiga toxin-producing E. coli O157, and therefore, it may be a helpful tool in foodborne disease surveillance programs [74,75].…”

Section: Genome-based Methodsmentioning

confidence: 99%

Detection of microbial food contaminants and their products by capillary electromigration techniques

García‐Cañas

Cifuentes

2007

Electrophoresis

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Abbreviations: AFLP, amplified fragment length polymorphism; AGE, agarose gel electrophoresis, ASP, amnesic shellfish poisoning; BGE, background electrolyte; CFP, ciguatera fish poisoning; DA, domoic acid; DCIP, 2,6-dichlorophenolindophenol; DSP, diarrhetic shellfish poisoning; DTX, dinophysistoxin; ERIC, enterobacterial repetitive intergenic consensus; FB1, Fumonisin type 1; GTX, gonyautoxins; MC, microcystins; MLVA, multiple locus variable-number tandem-repeat; MTX, maitotoxin; NEO, neosaxitosin; NSP, neurotoxic shellfish poisoning; OA, ochratoxin A; OkA, okadaic acid; PSP, paralytic shellfish poisoning; PEO, polyethylenoxide; STX, saxitoxin; TMV, tobacco mosaic virus; T-RFLP, terminal restriction fragment length polymorphism; TTX, tetrodotoxin; UTIs, urinary tract infections.

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Multiple-locus variable-number tandem-repeats analysis of Salmonella enterica subsp. enterica serovar Typhimurium using PCR multiplexing and multicolor capillary electrophoresis

Cited by 252 publications

References 4 publications

Vibrio cholerae Strain Typing and Phylogeny Study Based on Simple Sequence Repeats

Vibrio cholerae Strain Typing and Phylogeny Study Based on Simple Sequence Repeats

Genotypic diversity, pathogenic potential and the resistance profile of Salmonella Typhimurium strains isolated from humans and food from 1983 to 2013 in Brazil

Detection of microbial food contaminants and their products by capillary electromigration techniques

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