Multiple-locus variable-number tandem-repeats analysis of Listeria monocytogenes using multicolour capillary electrophoresis and comparison with pulsed-field gel electrophoresis typing

Lindstedt, Bjørn-Arne; Tham, Wilhelm; Danielsson-Tham, Marie-Louise; Vardund, Traute; Helmersson, Seved; Kapperud, Georg

doi:10.1016/j.mimet.2007.11.012

Cited by 64 publications

(51 citation statements)

References 22 publications

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“…First, we included all 15 of the VNTR loci described previously (25)(26)(27)(28)(29). The VNTR loci common to more than one of the above studies were amplified with all of the different PCR primer sets defined previously (Table 1).…”

Section: Bacterial Isolatesmentioning

confidence: 99%

“…MLVA relies on the study of the variability of the number of tandem repeats at specific loci in bacterial genomes. MLVA schemes are constructed on the basis of an open choice of several VNTR chromosomal loci, and five different MLVA schemes have been developed almost simultaneously for L. monocytogenes strain typing (25)(26)(27)(28)(29). Subsequent use of MLVA in outbreak investigations and strain diversity studies has relied on these different schemes or combinations thereof (30)(31)(32)(33)(34).…”

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confidence: 99%

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Optimized Multilocus Variable-Number Tandem-Repeat Analysis Assay and Its Complementarity with Pulsed-Field Gel Electrophoresis and Multilocus Sequence Typing for Listeria monocytogenes Clone Identification and Surveillance

et al. 2013

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iPopulations of the food-borne pathogen Listeria monocytogenes are genetically structured into a small number of major clonal groups, some of which have been implicated in multiple outbreaks. The goal of this study was to develop and evaluate an optimized multilocus variable number of tandem repeat (VNTR) analysis (MLVA) subtyping scheme for strain discrimination and clonal group identification. We evaluated 18 VNTR loci and combined the 11 best ones into two multiplexed PCR assays (MLVA-11). A collection of 255 isolates representing the diversity of clonal groups within phylogenetic lineages I and II, including representatives of epidemic clones, were analyzed by MLVA-11, multilocus sequence typing (MLST), and pulsed-field gel electrophoresis (PFGE). MLVA-11 had less discriminatory power than PFGE, except for some clones, and was unable to distinguish some epidemiologically unrelated isolates. Yet it distinguished all major MLST clones and therefore constitutes a rapid method to identify epidemiologically relevant clonal groups. Given its high reproducibility and high throughput, MLVA represents a very attractive first-line screening method to alleviate the PFGE workload in outbreak investigations and listeriosis surveillance.

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Section: Bacterial Isolatesmentioning

confidence: 99%

mentioning

confidence: 99%

Optimized Multilocus Variable-Number Tandem-Repeat Analysis Assay and Its Complementarity with Pulsed-Field Gel Electrophoresis and Multilocus Sequence Typing for Listeria monocytogenes Clone Identification and Surveillance

et al. 2013

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“…To help resolve such scenarios, multiple-locus variable-number tandem-repeat analysis (MLVA), which exploits the patterns of amplicons generated by primers that flank variable-length regions of the genome, is sometimes employed (11,12). However, as with PFGE, isolates of different origins can have identical MLVA patterns, because this technique will not resolve single nucleotide differences.…”

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confidence: 99%

Precise Dissection of an Escherichia coli O157:H7 Outbreak by Single Nucleotide Polymorphism Analysis

et al. 2013

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The current pathogen-typing methods have suboptimal sensitivities and specificities. DNA sequencing offers an opportunity to type pathogens with greater degrees of discrimination using single nucleotide polymorphisms (SNPs) than with pulsed-field gel electrophoresis (PFGE) and other methodologies. In a recent cluster of Escherichia coli O157:H7 infections attributed to salad bar exposures and romaine lettuce, a subset of cases denied exposure to either source, although PFGE and multiple-locus variable-number tandem-repeat analysis (MLVA) suggested that all isolates had the same recent progenitor. Interrogation of a preselected set of 3,442,673 nucleotides in backbone open reading frames (ORFs) identified only 1 or 2 single nucleotide differences in 3 of 12 isolates from the cases who denied exposure. The backbone DNAs of 9 of 9 and 3 of 3 cases who reported or were unsure about exposure, respectively, were isogenic. Backbone ORF SNP set sequencing offers pathogen differentiation capabilities that exceed those of PFGE and MLVA.

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“…Out of the 16 VNTRs published, only eight Lm-3, Lm-8, Lm-10, Lm-11, Lm-15, JLR4, LMV6-JLR and LMV9-JLR were selected here because (1) their repeat length was greater than or equal to 9 bp as demonstrated on a large panel of human and food strains (Sperry et al, 2008;Larson et al 2010;Lindstedt et al, 2008;Murphy et al, 2007). For six out of eight loci (Lm-3, Lm-10, Lm-11, Lm-15, LMV6-JLR and LMV9-JLR), the size of the amplification products observed on the agarose gels differed between the runs.…”

Section: Discussionmentioning

confidence: 99%

The Use of Pulsed Field Gel Electrophoresis in Listeria monocytogenes Sub-Typing - Comparison with MLVA Method Coupled with Gel Electrophoresis

Roussel¹,

Vignaud²,

Larsson³

et al. 2012

Gel Electrophoresis - Principles and Basics

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Multiple-locus variable-number tandem-repeats analysis of Listeria monocytogenes using multicolour capillary electrophoresis and comparison with pulsed-field gel electrophoresis typing

Cited by 64 publications

References 22 publications

Optimized Multilocus Variable-Number Tandem-Repeat Analysis Assay and Its Complementarity with Pulsed-Field Gel Electrophoresis and Multilocus Sequence Typing for Listeria monocytogenes Clone Identification and Surveillance

Optimized Multilocus Variable-Number Tandem-Repeat Analysis Assay and Its Complementarity with Pulsed-Field Gel Electrophoresis and Multilocus Sequence Typing for Listeria monocytogenes Clone Identification and Surveillance

Precise Dissection of an Escherichia coli O157:H7 Outbreak by Single Nucleotide Polymorphism Analysis

The Use of Pulsed Field Gel Electrophoresis in Listeria monocytogenes Sub-Typing - Comparison with MLVA Method Coupled with Gel Electrophoresis

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