Identification and Differentiation of Species and Strains of Arthrobacter and Microbacterium barkeri Isolated from Smear Cheeses with Amplified Ribosmal DNA Restriction Analysis (ARDRA) and Pulsed Field Gel Electrophoresis (PFGE)

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.

Download Full-text

Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing.

Journal of Clinical Microbiology ◽

10.1128/jcm.33.9.2233-2239.1995 ◽

1995 ◽

Vol 33 (9) ◽

pp. 2233-2239 ◽

Cited By ~ 5046

Author(s):

F C Tenover ◽

R D Arbeit ◽

R V Goering ◽

P A Mickelsen ◽

B E Murray ◽

...

Keyword(s):

Bacterial Strain ◽

Gel Electrophoresis ◽

Pulsed Field Gel Electrophoresis ◽

Strain Typing ◽

Chromosomal Dna ◽

Restriction Patterns ◽

Pulsed Field ◽

Dna Restriction

Download Full-text

Listeria monocytogenes in Five Sardinian Swine Slaughterhouses: Prevalence, Serotype, and Genotype Characterization

Journal of Food Protection ◽

10.4315/0362-028x.jfp-12-505 ◽

2013 ◽

Vol 76 (11) ◽

pp. 1863-1867 ◽

Cited By ~ 15

Author(s):

DOMENICO MELONI ◽

FRANCESCA PIRAS ◽

ANNA MUREDDU ◽

FEDERICA FOIS ◽

SIMONETTA GIANNA CONSOLATI ◽

...

Keyword(s):

Listeria Monocytogenes ◽

Gel Electrophoresis ◽

Restriction Analysis ◽

Pulsed Field Gel Electrophoresis ◽

Pork Meat ◽

Pulsed Field ◽

Restriction Profiles ◽

Full Complement ◽

Contact Surfaces ◽

Slaughtered Pigs

In a 3-year study (2008 to 2011) to estimate the prevalence and the contamination sources of Listeria monocytogenes in pork meat in Sardinia, Italy, 211 samples were collected from five Sardinian swine slaughterhouses: 171 samples from slaughtered pigs and 40 from the slaughterhouse environment. Fifty L. monocytogenes isolates were characterized by PCR-based serotyping, presence of virulence-associated genes, and pulsed-field gel electrophoresis restriction analysis. The overall prevalence of L. monocytogenes was 33% in swine carcasses, 7% in cecal material, 23% on meat contact surfaces, and 25% on noncontact surfaces. Only two serotypes were detected: 1/2c (78%) and 1/2a (22%). In all, based on the presence of virulence-associated genes, eight pathogenic profiles were detected. Only 42% of all isolates carried the full complement of virulence-associated genes and were allotted to profile 1. Six pulsed-field gel electrophoresis profiles persisted in the slaughterhouses; restriction profiles appeared to be specific to each plant.

Download Full-text

DNA restriction patterns produced by pulsed-field gel electrophoresis in Moraxella catarrhalis isolated from different geographical areas

Epidemiology and Infection ◽

10.1017/s0950268899002381 ◽

1999 ◽

Vol 122 (3) ◽

pp. 417-422 ◽

Cited By ~ 9

Author(s):

G. MARTINEZ ◽

K. AHMED ◽

C. H. ZHENG ◽

K. WATANABE ◽

K. OISHI ◽

...

Keyword(s):

Gel Electrophoresis ◽

Moraxella Catarrhalis ◽

Genomic Dna ◽

Respiratory Infections ◽

Pulsed Field Gel Electrophoresis ◽

Dynamic Changes ◽

Restriction Patterns ◽

Pulsed Field ◽

Time Period ◽

Dna Restriction

Pulsed field gel electrophoresis (PFGE) of the genomic DNA of Moraxella catarrhalis was done in 172 strains isolated from sputum of patients with respiratory infections in Nagasaki (130 strains), Europe (14 strains), Thailand (6 strains), Uganda (3 strains), Bangladesh (5 strains) and Kuwait (14 strains). Restriction endonuclease with SmaI generated 4–16 DNA fragments ranging from 1000 kb to 24·25 kb and was classified into 31 major groups. It was found that there were wide variations of DNA restriction patterns of strains isolated from the same and different geographical areas. DNA restriction patterns of strains isolated in Nagasaki during the last 12 years showed dynamic changes of the predominant strains in each time period. We conclude from this study that PFGE is a suitable method to document interstrain variation in M. catarrhalis.

Download Full-text

Comparative Analysis of Infrequent-Restriction-Site PCR and Pulsed-Field Gel Electrophoresis for Epidemiological Typing ofLegionella pneumophila Serogroup 1 Strains

Journal of Clinical Microbiology ◽

10.1128/jcm.36.1.161-167.1998 ◽

1998 ◽

Vol 36 (1) ◽

pp. 161-167 ◽

Cited By ~ 33

Author(s):

Serge Riffard ◽

François Lo Presti ◽

François Vandenesch ◽

Françoise Forey ◽

Monique Reyrolle ◽

...

Keyword(s):

Legionella Pneumophila ◽

Gel Electrophoresis ◽

Restriction Site ◽

Pulsed Field Gel Electrophoresis ◽

Pfge Analysis ◽

Pcr Assay ◽

Pulsed Field ◽

Restriction Sites ◽

Dna Restriction ◽

High Level

Two methods were compared for the analysis of 48 unrelated and epidemiologically related Legionella pneumophila serogroup 1 isolates. These are the infrequent-restriction-site PCR (IRS-PCR) assay with adapters designed for XbaI and PstI restriction sites and the pulsed-field gel electrophoresis (PFGE) analysis determined after DNA restriction with SfiI. Both methods demonstrated a high level of discrimination with a similar capacity for differentiating 23 of the 24 unrelated isolates. PFGE analysis and IRS-PCR assay were both able to identify epidemiologically related isolates of L. pneumophila from three outbreaks. Hence, IRS-PCR assay appears to be a reproducible (intergel reproducibility, 100%) and discriminative (discriminatory index, ≥0.996) tool for typing of Legionella. Compared to PFGE, however, IRS-PCR presented an advantage through ease of performance and with attributes of rapidity and sensitivity of target DNA.

Download Full-text

Evaluation of PFGE and mtDNA restriction analysis methods to detect genetic diversity of saccharomyces cerevisiae strains associated to vitis vinifera

OENO One ◽

10.20870/oeno-one.2007.41.3.848 ◽

2007 ◽

Vol 41 (3) ◽

pp. 155

Author(s):

Mar Vilanova ◽

Sol Zamuz ◽

Antón Masa ◽

Carmen Sieiro

Keyword(s):

Saccharomyces Cerevisiae ◽

Genetic Diversity ◽

Mitochondrial Dna ◽

Vitis Vinifera ◽

Gel Electrophoresis ◽

Restriction Analysis ◽

Molecular Genetic ◽

Pulsed Field Gel Electrophoresis ◽

Pulsed Field ◽

Mtdna Restriction Analysis

Aims: The aim of this work was realize a comparative study of two different methods of Saccharomycec cerevisiae yeast strain characterization.Methods and results: Pulsed-field gel electrophoresis analysis (PFGE) and mitochondrial DNA (mtDNA) restriction analysis have been carried out to differentiate strains of Saccharomyces cerevisiae associated to Vitis vinifera musts from different Galicia wineyard (NW Spain). Seventeen strains isolated from wineries from Galicia were used in this study.Conclusion: The results have showed that although PFGE analysis technique has greater discriminatory power than mtDNA restriction analysis to detect genetic diversity in Saccharomyces cerevisiae, some clones with the same PFGE profile can only be differentiated by mtDNA restriction analysis.Significance and impact of study: Pulsed-field gel electrophoresis analysis of chromosome (PFGE), by its discriminating power, constitute an ideal technique for the differentiation of Saccharomyces cerevisiae strains in biotechnological industries, however, mitochondrial DNA (mtDNA) restriction analysis is a rapid, simple and less expensive and time-consuming method. The results obtained demonstrate the value of using molecular genetic methods in taxonomic and ecological surveys.

Download Full-text