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 Relatedness of Listeria monocytogenes Isolates Recovered from Selected Ready-To-Eat Foods and Listeriosis Patients in the United States

2005 ◽  
Vol 71 (12) ◽  
pp. 8115-8122 ◽  
Author(s):  
Stefanie Evans Gilbreth ◽  
Jeff E. Call ◽  
F. Morgan Wallace ◽  
Virginia N. Scott ◽  
Yuhuan Chen ◽  
...  
Keyword(s):  
United States ◽  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Food Products ◽  
The United States ◽  
Pulsed Field Gel Electrophoresis ◽  
Clinical Isolates ◽  
Pulsed Field ◽  
Serotype 1 ◽  
Serotype 4B

ABSTRACT Pulsed-field gel electrophoresis and serotyping were performed for 544 isolates of Listeria monocytogenes, including 502 isolates recovered from contaminated samples from 31,705 retail ready-to-eat (RTE) food products and 42 isolates recovered from human cases of listeriosis. The isolates were from Maryland (294 isolates) and California (250 isolates) and were collected in 2000 and 2001. The isolates were placed into 16 AscI pulsogroups (level of relatedness within each group, ≥66%), 139 AscI pulsotypes (levels of relatedness, ≥25% to 100%), and eight serotypes (serotypes 1/2a, 1/2b, 1/2c, 3a, 3b, 4b, 4c, and 4d). The most frequently found pulsotypes belonged to either pulsogroup A (150 food isolates plus 4 clinical isolates) or pulsogroup B (104 food isolates plus 5 clinical isolates). The majority of the 502 food isolates were either serotype 1/2a (298 isolates) or serotype 1/2b (133 isolates), whereas the majority of the 42 clinical isolates were either serotype 1/2a (19 isolates) or serotype 4b (15 isolates). Additionally, 13 clinical isolates displayed pulsotypes also found in food isolates, whereas the remaining 29 clinical isolates displayed 24 unique pulsotypes. These data indicate that most (86%) of the L. monocytogenes subtypes found in the RTE foods sampled belonged to only two serotypes and that 90% of the isolates displayed 73 pulsotypes, with 107 isolates displaying pulsotype 1. These data should help define the distribution and relatedness of isolates found in RTE foods in comparison with isolates that cause listeriosis.

scholarly journals Comparison of Virulence Gene Identification, Ribosomal Spacer PCR, and Pulsed-Field Gel Electrophoresis for Typing of Staphylococcus aureus Strains Isolated from Cases of Subclinical Bovine Mastitis in the United States

2016 ◽  
Vol 54 (7) ◽  
pp. 1871-1876 ◽  
Author(s):  
Pamela R. F. Adkins ◽  
John R. Middleton ◽  
Lawrence K. Fox
Keyword(s):  
United States ◽  
Staphylococcus Aureus ◽  
Gel Electrophoresis ◽  
Virulence Gene ◽  
The United States ◽  
Pulsed Field Gel Electrophoresis ◽  
Gene Identification ◽  
Content Type ◽  
Pulsed Field

Staphylococcus aureusis one of the most important pathogens causing contagious mastitis in dairy cattle worldwide. The objectives of this study were to determine if recently describedS. aureusgenotype B was present among previously characterized isolates from cases of bovine intramammary infection in the United States and to compare pulsed-field gel electrophoresis (PFGE) to the combination of ribosomal spacer PCR (RS-PCR) and virulence gene identification for typing ofS. aureusstrains. The hypothesis was that isolates that were previously characterized as contagious would be identified as genotype B and that the results of the two strain-typing methods would be comparable. Isolates were selected from a collection ofS. aureusisolates from eight dairy farms. Mammary quarter milk somatic cell count (SCC) andN-acetyl-β-d-gluconaminidase (NAGase) activity data were known and used to evaluate strain pathogenicity. RS-PCR was performed with conventional gel electrophoresis, and PCR was used for toxin gene identification. RS-PCR patterns were associated with a specific virulence gene pattern, as previously reported. Five RS-PCR banding patterns were identified. None of the isolates were characterized as genotype B. No association between RS-PCR types and milk SCC was found; however, NAGase activity was significantly higher in milk from mammary glands infected with RS-PCR banding type 1 (RSP type 1) than in milk from those infected with RSP type 2. The discriminatory power values were 1.0 and 0.46 for PFGE and RS-PCR, respectively. These data suggest that genotype B may have a limited geographic distribution and that PFGE is more discriminatory than RS-PCR performed with conventional gel electrophoresis for typing ofS. aureusisolates of bovine origin.

scholarly journals Pulsed-Field Gel Electrophoresis Analysis of Vibrio vulnificus Strains Isolated from Taiwan and the United States

2004 ◽  
Vol 70 (9) ◽  
pp. 5153-5158 ◽  
Author(s):  
Hin-chung Wong ◽  
Shau-Yan Chen ◽  
Meng-Yi Chen ◽  
James D. Oliver ◽  
Lien-I Hor ◽  
...  
Keyword(s):  
United States ◽  
Gel Electrophoresis ◽  
Vibrio Vulnificus ◽  
The United States ◽  
Pulsed Field Gel Electrophoresis ◽  
Intraspecific Diversity ◽  
Pulsed Field ◽  
Clinical Strains ◽  
Discriminative Method ◽  
Geographic Regions

ABSTRACT Vibrio vulnificus is a marine bacterium that causes human wound infections and septicemia with a high mortality rate. V. vulnificus strains from different clinical and environmental sources or geographic regions have been successfully characterized by ribotyping and several other methods. Pulsed-field gel electrophoresis (PFGE) is a highly discriminative method, but previous studies suggested that it was not suitable for examining the correlation of V. vulnificus strains from different origins. We employed PFGE to determine its efficacy for characterizing V. vulnificus strains from different geographic regions, characterizing a total of 153 strains from clinical and environmental origins from the United States and Taiwan after SfiI or NotI digestion. V. vulnificus strains showed a high intraspecific diversity by PFGE after SfiI or NotI digestion, and about 12% of the strains could not be typed by the use of either of these enzymes. For PFGE with SfiI digestion, most of the clinical and environmental strains from the United States were grouped into cluster A, while the strains from Taiwan were grouped into other clusters. Clinical strains from the United States showed a higher level of genetic homogeneity than clinical strains from Taiwan, and environmental strains from both regions showed a similarly high level of heterogeneity. PFGE with NotI digestion was useful for studying the correlation of clinical strains from the United States and Taiwan, but it was not suitable for analyzing environmental strains. The results showed that PFGE with SfiI digestion may be used to characterize V. vulnificus strains from distant geographic regions, with NotI being a recommended alternative enzyme.

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

1999 ◽  
Vol 122 (3) ◽  
pp. 417-422 ◽  
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.

Characteristics of Salmonella Bareilly isolated from commercial layer farms and raw shell eggs in Korea

2019 ◽  
Vol 99 (2) ◽  
pp. 425-427
Author(s):  
Kwang Won Seo ◽  
Min Chan Im ◽  
Yeong Bin Kim ◽  
Haan Woo Sung ◽  
Young Ju Lee
Keyword(s):  
United States ◽  
Antimicrobial Resistance ◽  
Gel Electrophoresis ◽  
Geographical Variation ◽  
The United States ◽  
Pulsed Field Gel Electrophoresis ◽  
Retail Markets ◽  
Pulsed Field ◽  
Shell Eggs ◽  
Epidemiological Characteristics

Salmonella enterica serovar Bareilly (S. Bareilly) has been among the top 20 most frequently isolated serovars in the United States and has been observed recently in layer flocks in Korea. Between 2013 and 2014, 45 S. Bareilly isolates were obtained from five commercial layer farms and nine retail markets in Korea. Among the 45 isolates, four pulsed-field gel electrophoresis patterns were observed, with pattern B being the predominant and comprising 67% of the 45 isolates. The most common antimicrobial resistance was streptomycin (24.4%) and cephalothin (6.7%). This is the first report describing epidemiological characteristics of S. Bareilly, including geographical variation, in Korea.

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