Use of pulsed field gel electrophoresis to compare large DNA-restriction fragments of Listeria monocytogenes strains belonging to serogroups 1/2 and 3

1991 ◽  
Vol 14 (3-4) ◽  
pp. 297-304 ◽  
Author(s):  
C. Buchrieser ◽  
R. Brosch ◽  
J. Rocourt
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Restriction Fragments ◽  
Pulsed Field ◽  
Dna Restriction Fragments ◽  
Dna Restriction

Pulsed-field gel electrophoresis applied for comparing Listeria monocytogenes strains involved in outbreaks

1993 ◽  
Vol 39 (4) ◽  
pp. 395-401 ◽  
Author(s):  
C. Buchrieser ◽  
R. Brosch ◽  
B. Catimel ◽  
J. Rocourt
Keyword(s):  
United States ◽  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
The United States ◽  
Pulsed Field Gel Electrophoresis ◽  
Chromosomal Dna ◽  
Restriction Patterns ◽  
Pulsed Field ◽  
Dna Restriction Fragments ◽  
Dna Restriction

Recent food-borne outbreaks of human listeriosis as well as numerous sporadic cases have been mainly caused by Listeria monocytogenes serovar 4b strains. Thus, it was of interest to find out whether a certain clone or a certain few clones were responsible for these cases and especially for outbreaks., We used pulsed-field gel electrophoresis of large chromosomal DNA restriction fragments generated by ApaI, SmaI, or NotI to analyse 75 L. monocytogenes strains isolated during six major and eight smaller recent listeriosis outbreaks. These strains could be divided into 20 different genomic varieties. Thirteen of 14 strains isolated during major epidemics in Switzerland (1983–1987), the United States (California, 1985) and Denmark (1985–1987) demonstrated indistinguishable DNA restriction patterns. In contrast, strains responsible for the outbreaks in Canada (Nova Scotia, 1981), the United States (Massachusetts, 1983), France (Anjou, 1975–1976), New Zealand (1969), and Austria (1986) and some smaller outbreaks in France (1987, 1988, 1989) were each characterized by particular combinations of DNA restriction patterns. Seventy-seven percent of the tested strains could be classified into the previously described ApaI group A (Brosch et al. 1991), demonstrating a very close genomic relatedness. Because 49% of the epidemic strains selected for this study belonged to phagovar 2389/2425/3274/2671/47/108/340 or 2389/47/108/340, fifty-six additional strains of these phagovars, isolated from various origins, were also typed to determine whether differences in DNA restriction profiles between epidemic and randomly selected strains of the same phagovars could be pointed out. Variations in DNA patterns appeared more frequently within randomly selected strains than within epidemic strains.Key words: Listeria monocytogenes, listeriosis, typing, pulsed-field gel electrophoresis, epidemic.

scholarly journals Caracterização molecular de Listeria monocytogenes oriundas de cortes cárneos bovinos e de abatedouros frigoríficos de bovinos localizados no Distrito Federal, Brasil

2016 ◽  
Vol 36 (10) ◽  
pp. 957-964 ◽  
Author(s):  
Joana M. Palma ◽  
◽  
Rodrigo C. Lisboa ◽  
Dália P. Rodrigues ◽  
André F.M. Santos ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Distrito Federal

RESUMO: Este trabalho teve como objetivo realizar a detecção de cepas de Listeria monocytogenes de cortes cárneos bovinos bem como no ambiente de abatedouros frigoríficos localizados no Distrito Federal, promover a sorotipificação pela reação em cadeia da polimerase (PCR), realizar antibiograma e submeter às cepas à eletroforese de campo pulsado (Pulsed-field gel electrophoresis - PFGE). Foram analisados um total de 125 cortes cárneos bovinos, 45 amostras de swabs de carcaças e 43 amostras de swabs em que foram detectados 13 cepas de Listeria monocytogenes, sendo 11 em cortes cárneos bovinos e 2 swabs de ambiente em um abatedouro frigorifico. Não foram isoladas cepas de swabs de carcaça. Dentre as 13 cepas de Listeria monocytogenes foram encontradas seis cepas do sorotipo 4b, cinco do sorotipo 1/2c e duas cepas do sorotipo 1/2a. Dentre as 11 cepas de L. monocytogenes encontradas em cortes cárneos bovino, uma (9,1%) cepa apresentou resistência a eritromicina, outra (9,1%) cepa a gentamicina e outra a ciprofloxacina (9,1%) e todas as cepas (100%) apresentaram resistência ao Ác. Nalidíxico. Das duas (2) cepas oriundas de ralos de abatedouro frigorífico, todas (100%) apresentaram resistência ao Ác. Nalidíxico e a sulfonamidas. A análise por eletroforese de campo pulsante (PFGE) demonstrou 13 diferentes pulsotipos, em que foram agrupados em 3 diferentes grupos clonais, que coincidentemente se correlacionavam com os 3 diferentes sorotipos encontrados sugerindo uma ampla disseminação desses perfis no Distrito Federal.

Isolation and Pulsed-Field Gel Electrophoresis Typing of Listeria monocytogenes from Modified Atmosphere Packaged Fresh-Cut Vegetables Collected in Ireland

2006 ◽  
Vol 69 (10) ◽  
pp. 2524-2528 ◽  
Author(s):  
GILLIAN A. FRANCIS ◽  
DAVID O'BEIRNE
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Fruits And Vegetables ◽  
Pulsed Field Gel Electrophoresis ◽  
Leafy Vegetables ◽  
Modified Atmosphere ◽  
Pulsed Field ◽  
Fruit Samples ◽  
Fresh Cut ◽  
Pfge Profiles

The incidence of Listeria monocytogenes in modified atmosphere packaged fresh-cut fruits and vegetables from chill cabinets of a supermarket in Ireland was investigated over a 2-year period. Overall, 9.58% of fresh-cut produce was contaminated with Listeria spp. Various species of Listeria were isolated from samples, including L. monocytogenes, L. seeligeri, L. innocua, L. welshimeri, and L. ivanovii. No fruit samples contained detectable L. monocytogenes. Overall, a total of 21 L. monocytogenes isolates (2.9% of samples) were recovered from a range of products, including dry coleslaw mix (80% shredded cabbage and 20% shredded carrot), bean sprouts, and leafy vegetables such iceberg, romaine, and radicchio lettuce and mixed salad leaves (curly endive, escarole, and radicchio leaves). Dry coleslaw mix appeared to have the highest incidence of Listeria contamination (20%) compared with other products. Listeria contamination was more frequent (P < 0.05) during the summer and autumn months than during the winter and spring months. The 21 L. monocytogenes isolates were subsequently subtyped by genomic macrorestriction techniques using ApaI with pulsed-field gel electrophoresis (PFGE). PFGE of digested DNA produced bands of 79 to 518 kb. Four PFGE profiles were identified, and approximately 50% of the isolates were associated with profile 1. This study indicates that fresh-cut vegetables packaged under a modified atmosphere can support growth of numerous species of Listeria, including L. monocytogenes.

scholarly journals Sources of Listeria monocytogenes Contamination in a Cold-Smoked Rainbow Trout Processing Plant Detected by Pulsed-Field Gel Electrophoresis Typing

1999 ◽  
Vol 65 (1) ◽  
pp. 150-155 ◽  
Author(s):  
Tiina Autio ◽  
Sebastian Hielm ◽  
Maria Miettinen ◽  
Anna-Maija Sjöberg ◽  
Kaarina Aarnisalo ◽  
...  
Keyword(s):  
Rainbow Trout ◽  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Hot Water ◽  
Processing Plant ◽  
Pulsed Field ◽  
Eradication Program ◽  
Fish Samples ◽  
Raw Fish

ABSTRACT Sites of Listeria monocytogenes contamination in a cold-smoked rainbow trout (Oncorhynchus mykiss) processing plant were detected by sampling the production line, environment, and fish at different production stages. Two lots were monitored. The frequency of raw fish samples containing L. monocytogenes was low. During processing, the frequency of fish contaminated with L. monocytogenes clearly rose after brining, and the most contaminated sites of the processing plant were the brining and postbrining areas. A total of 303 isolates from the raw fish, product, and the environment were characterized by pulsed-field gel electrophoresis (PFGE). PFGE yielded nine pulsotypes, which formed four clusters. The predominating L. monocytogenespulsotypes of the final product were associated with brining and slicing, whereas contaminants of raw fish were not detected in the final product. Air-mediated contamination in the plant could not be proved. In accordance with these results, an L. monocytogenes eradication program was planned. The use of hot steam, hot air, and hot water seemed to be useful in eliminatingL. monocytogenes. None of the control samples taken in the 5 months after the eradication program was implemented containedL. monocytogenes.

scholarly journals Use of pulsed field gel electrophoresis and transposon mutagenesis to estimate the minimal number of genes required for motility in Caulobacter crescentus.

Genetics ◽  
1989 ◽  
Vol 123 (4) ◽  
pp. 649-654 ◽  
Author(s):  
B Ely ◽  
T W Ely
Keyword(s):  
Gel Electrophoresis ◽  
Caulobacter Crescentus ◽  
Pulsed Field Gel Electrophoresis ◽  
Restriction Fragments ◽  
Pulsed Field ◽  
Transposon Insertion ◽  
Number Of Genes ◽  
Physical And Genetic Map ◽  
Insertion Mutations ◽  
Flagellar Genes

Abstract To facilitate the mapping of transposon insertion mutations in Caulobacter crescentus, we have used pulsed field gel electrophoresis to construct a detailed physical and genetic map of the C. crescentus genome. Restriction fragments were generated by DraI, AseI, or SpeI which cleave the C. crescentus 40, 13, and 26 times, respectively, and Tn5 insertions were used to align the restriction fragments generated by each of the enzymes. The utility of the resulting map was demonstrated by determining the chromosomal locations of a collection of flagellar mutations. As a result of this study, we were able to identify ten new flagellar genes at various locations on the chromosome. Thus, at least 48 genes are required for the assembly of a functional flagellum in C. crescentus.

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