Comparison of repetitive element sequence-based PCR with multilocus enzyme electrophoresis and pulsed field gel electrophoresis for typing Listeria monocytogenes food isolates

Harvey, J.; Norwood, D. E.; Gilmour, A.

doi:10.1016/j.fm.2003.08.003

Cited by 19 publications

(14 citation statements)

References 19 publications

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“…Despite the high number of papers published on the successful application of rep-PCR on Listeria isolated from foods (e.g., Jersek et al, 1999;Harvey et al, 2004;Zunabovic et al, 2012) typing data on silage collected samples lack in literature, and the presented resulted were difficult to compare. However, the present data are in agreement with those reported by an earlier study using ribotyping (Ryser et al, 1997).…”

Section: Discussionmentioning

confidence: 99%

“…Dairy cows may be directly exposed to L. monocytogenes through the ingestion of improperly fermented silage (pH >5.0) contaminated before ensiling (Fenlon, 1988), and L. monocytogenes may then reach bulk tanks as a result of fecal contamination during milking, as also reported for spore-forming bacteria (Vissers et al, 2007). Other than Listeria detection, currently strain typing has largely been applied to explore subtype frequency and distribution: some authors applied repetitive element sequence-based PCR (REP) for characterizing isolates collected from dairy primary production as well as from the food processing environment (Harvey et al, 2004;Van Kessel et al, 2005;Chou and Wang, 2006), indicating the putative transmission/contamination paths.…”

Section: Introductionmentioning

confidence: 97%

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Detection, identification, and typing of Listeria species from baled silages fed to dairy cows

Nucera

Grassi

Morra

et al. 2016

Journal of Dairy Science

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Anaerobiosis, critical for successful ensilage, constitutes a challenge in baled silages. The loss of complete anaerobiosis causes aerobic deterioration and silages undergo dry matter and nutrient losses, pathogen growth, and mycotoxin production. Silage may represent an ideal substrate for Listeria monocytogenes, a pathogen of primary concern in several cheeses. The aim of this research was to investigate the occurrence of Listeria in baled silage fed to cows producing milk for a protected designation of origin cheese, and to characterize isolates by repetitive sequence-based PCR. Listeria spp. were detected in 21 silages and L. monocytogenes in 6 out of 80 of the analyzed silages; 67% of positives were found in molded zones. Results of the PCR typing showed genotypic homogeneity: 72.9 and 78.8% similarity between strains of Listeria spp. (n = 56) and L. monocytogenes (n = 24), respectively. Identical profiles were recovered in molded and nonmolded areas, indicating that contamination may have occurred during production. The application of PCR allowed the unambiguous identification of Listeria isolated from baled silages, and repetitive sequence-based PCR allowed a rapid and effective typing of isolates. Results disclose the potential of the systematic typing of Listeria in primary production, which is needed for the understanding of its transmission pathways.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

Detection, identification, and typing of Listeria species from baled silages fed to dairy cows

Nucera

Grassi

Morra

et al. 2016

Journal of Dairy Science

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“…Rep-PCR uses primers that target noncoding short repetitive sequence elements interspersed throughout the genome, whose function is probably associated with gene regulation (Harvey et al, 2004). The amplified DNA fragments, when separated by electrophoresis, constitute a genomic fingerprint that can be employed for bacterial discrimination.…”

Section: Introductionmentioning

confidence: 99%

Semi-Automated Repetitive-Sequence-Based Polymerase Chain Reaction Compared to Pulsed-Field Gel Electrophoresis forListeria monocytogenesSubtyping

Roussel

Félix²,

Colaneri³

et al. 2010

Foodborne Pathogens and Disease

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Listeriosis is a severe infection that mainly affects pregnant women, neonates, and immuno-compromised adults. The commercially available semi-automated repetitive-sequence-based polymerase chain reaction assay system, DiversiLab, has been successfully used for subtyping several species of bacteria. In this article we compare the DiversiLab System with macrorestriction analysis by pulsed-field gel electrophoresis (PFGE), which is currently the gold standard for molecular subtyping of Listeria monocytogenes. We used a panel of 116 human and food L. monocytogenes isolates for the comparative evaluation. Among these isolates, there were 4 pairs of duplicates, 13 strains were epidemiologically related, and the remaining food isolates were epidemiologically unrelated. The isolates of different serotypes represented distinct DiversiLab types (DTs) and ApaI/AscI-PFGE types except for one DT-containing isolates of two serotypes, 4b and 1/2b. The four duplicates displayed the same DT and ApaI/AscI PFGE type demonstrating the good reproducibility of the two methods. The epidemiologically related strains were clustered in the same DT and PFGE type. The Simpson's index of diversity was 0.954; 0.988; 0.994; and 0.998 for DiversiLab, AscI-PFGE, ApaI-PFGE, and AscI/ApaI-PFGE, respectively. Thus, PFGE was more discriminating than DiversiLab. However, for 1/2a serotype strains, six AscI-PFGE, three ApaI-PFGE, and one ApaI/AscI PFGE type were divided into different DTs. DiversiLab enabled a good discrimination between serotype 1/2a strains. DiversiLab is less labor intensive than PFGE and provides results in <24 hours compared with 30 hours to 3 days for PFGE from the time a pure culture of the bacteria has been obtained. On the basis of these results, DiversiLab may be useful for tracking the source of contamination in food-processing facilities and their environments. Also, DiversiLab may be more appropriate for long-term epidemiological studies where less discrimination is needed.

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“…(95 %) observed within each sample was used to define identical genotypes (Ravelo et al 2003). These parameters were accurately evaluated and set after performing repeatability assays for minimizing the intra-sample variation, i.e., faint bands, which may impair the epidemiologic interpretation of the results (Harvey 2004). However, the selected 95 % S.L.…”

Section: Resultsmentioning

confidence: 99%

Diagnostic Performance of rep-PCR as a Rapid Subtyping Method for Listeria monocytogenes

et al. 2012

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Previously pulsed field gel electrophoresistyped Listeria monocytogenes isolates (N095) were tested by repetitive element sequence-based PCRs (repPRCs). A combined rep-PCR typing approach showed 95 % repeatability, 0.98 discriminatory power, 95.5 % sensitivity, 75 % specificity, 91.2 % predictive positive value, and 85.7 % predictive negative value. Hence, rep-PCR represents an efficient and rapid subtyping method for L. monocytogenes.

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Comparison of repetitive element sequence-based PCR with multilocus enzyme electrophoresis and pulsed field gel electrophoresis for typing Listeria monocytogenes food isolates

Cited by 19 publications

References 19 publications

Detection, identification, and typing of Listeria species from baled silages fed to dairy cows

Detection, identification, and typing of Listeria species from baled silages fed to dairy cows

Semi-Automated Repetitive-Sequence-Based Polymerase Chain Reaction Compared to Pulsed-Field Gel Electrophoresis forListeria monocytogenesSubtyping

Diagnostic Performance of rep-PCR as a Rapid Subtyping Method for Listeria monocytogenes

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