Comparative study of five different techniques for epidemiological typing of Escherichia coli O157

Grif, Katharina; Karch, Helge; Schneider, Christian; Daschner, F; Beutin, Lothar; Cheasty, T.; Smith, H. R.; Rowe, B.; Dierich, M P; Allerberger, Franz

doi:10.1016/s0732-8893(98)00103-5

Cited by 65 publications

(56 citation statements)

References 31 publications

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“…PT2 and PT8 were predominant among human and bovine STEC O157:H7 strains in Spain as well as in many other European countries, including Belgium, Finland, Germany, Italy, England, and Scotland (20,22,23,30,33,34,46,58 . According to their phage types, stx genotypes, and phenotypes, the STEC O157:H7 strains isolated in Spain were very similar to those isolated in other parts of Europe.…”

Section: Discussionmentioning

confidence: 99%

Phage Types and Genotypes of Shiga Toxin-Producing Escherichia coli O157:H7 Isolates from Humans and Animals in Spain: Identification and Characterization of Two Predominating Phage Types (PT2 and PT8)

Mora

Blanco

et al. 2004

J Clin Microbiol

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Phage typing and DNA macrorestriction fragment analysis by pulsed-field electrophoresis (PFGE) were used for the epidemiological subtyping of a collection of Shiga toxin-producing Escherichia coli (STEC) O157:H7 strains isolated in Spain between 1980 and 1999. Phage typing distinguished a total of 18 phage types among 171 strains isolated from different sources (67 humans, 82 bovines, 12 ovines, and 10 beef products). However, five phage types, phage type 2 (PT2; 42 strains), PT8 (33 strains), PT14 (14 strains), PT21/28 (11 strains), and PT54 (16 strains), accounted for 68% of the study isolates. PT2 and PT8 were the most frequently found among strains from both humans (51%) and bovines (46%). Interestingly, we detected a significant association between PT2 and PT14 and the presence of acute pathologies. A group of 108 of the 171 strains were analyzed by PFGE, and 53 distinct XbaI macrorestriction patterns were identified, with 38 strains exhibiting unique PFGE patterns. In contrast, phage typing identified 15 different phage types. A total of 66 phage type-PFGE subtype combinations were identified among the 108 strains. PFGE subtyping differentiated between unrelated strains that exhibited the same phage type. The most common phage type-PFGE pattern combinations were PT2-PFGE type 1 (1 human and 11 bovine strains), PT8-PFGE type 8 (2 human, 6 bovine, and 1 beef product strains), PT2-PFGE subtype 4A (1 human, 3 bovine, and 1 beef product strains). Nine (29%) of 31 human strains showed phage type-PFGE pattern combinations that were detected among the bovine strains included in this study, and 26 (38%) of 68 bovine strains produced phage type-PFGE pattern combinations observed among human strains included in this study, confirming that cattle are a major reservoir of strains pathogenic for humans. PT2 and PT8 strains formed two groups which differed from each other in their motilities, stx genotypes, PFGE patterns, and the severity of the illnesses that they caused.

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Section: Discussionmentioning

confidence: 99%

Phage Types and Genotypes of Shiga Toxin-Producing Escherichia coli O157:H7 Isolates from Humans and Animals in Spain: Identification and Characterization of Two Predominating Phage Types (PT2 and PT8)

Mora

Blanco

et al. 2004

J Clin Microbiol

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“…In this study, six random primers were used (M13, 1247, 1254, 1253, 1281 and A04) and all of which had previously been utilized in E. coli population studies (Pacheco et al 1997, Grif et al 1998, Regua-Mangia et al 2004). After initial amplification reactions, three of them (A04, 1254 and M13) were selected for RAPD typing since their profiles presented reproducibility, higher discriminatory power and distinct polymorphic bands.…”

Section: Resultsmentioning

confidence: 99%

Genetic relationship between Escherichia coli strains isolated from dairy mastitis and from the stable fly Stomoxys calcitrans

et al. 2016

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The stable fly Stomoxys calcitrans (Linnaeus, 1758) has been described as a potential spreader of infectious agents to cattle herds. Among the agents transmitted by this fly, Escherichia coli has attracted attention due to its potential to cause gastrointestinal disorders as well as environmental mastitis in dairy cows. Therefore, the aim of this study was to isolate and to assess the genetic diversity and the clonal relatedness among E. coli isolates from the milk of dairy mastitis and from stable flies anatomical sites by the Random Amplification of Polymorphic DNA (RAPD-PCR) technique. The molecular typing revealed a high degree of genetic polymorphism suggesting that these microorganisms have a non-clonal origin. Identical electrophoretic profiles were observed between E. coli isolates from different flies, different mammary quarters of the same cow and from cows on a single farm. These results reveal the circulation of the same bacterial lineages and suggest the role of the stable fly in bacterial dispersion. Considering the high pathogenic potential of this bacterial species, our findings alert to a more effective health surveillance.INDEX TERMS: Random amplification of polymorphic DNA, RAPD, milk, stable fly, clonal species.

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“…PFGE was more discriminatory than both phage typing (1,29) and differentiation based on the presence of virulence genes (38). However, it is unclear when differences in PFGE profiles should be regarded as significant genetic differences (20). Tenover et al (40) proposed that for small (n ϭ Ͻ31) collections of microorganisms related to outbreaks of human disease, minor band differences should not be regarded as major genetic shifts and that only isolates differing by seven or more bands should be regarded as unrelated, while those differing by four to six bands should be regarded as possibly related.…”

Section: Figmentioning

confidence: 99%

“…Dalla-Costa et al (16), using separate BglI and HindIII digestions of one E. coli O157:HϪ, nine E. coli O157: H7, and three E. coli O157:H43 isolates, found that the 11 motile isolates had identical ribotypes, while the sole E. coli O157:HϪ isolate had another, closely related profile. Grif et al (20) used EcoRI and PvuII in separate digestions to automatically ribotype (riboprint) 47 E. coli O157 isolates originating from human patients, food (unrelated to the human cases), or healthy calves and produced five different ribotypes. In contrast, in the present study, 154 unrelated E. coli O157 isolates produced 46 different ribotypes.…”

Section: Figmentioning

confidence: 99%

“…Ribotyping has been used to genetically fingerprint some major human pathogens, including Salmonella enterica serovar Enteritidis (27), Campylobacter jejuni (21), and Listeria mono-cytogenes (32), among species of other genera. However, there are few reports on ribotyping of E. coli O157, and results have indicated that ribotyping is not a very discriminatory technique (16,20,30). Wachsmuth et al (41) found that ribotyping with NcoI produced maximum discrimination of E. coli O157:H7 and that it may have been more stable than plasmid profiling.…”

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confidence: 99%

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Combined Use of Two Genetic Fingerprinting Methods, Pulsed-Field Gel Electrophoresis and Ribotyping, for Characterization of Escherichia coli O157 Isolates from Food Animals, Retail Meats, and Cases of Human Disease

et al. 2002

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Two genetic fingerprinting techniques, pulsed-field gel electrophoresis (PFGE) and ribotyping, were used to characterize 207 Escherichia coli O157 isolates from food animals, foods of animal origin, and cases of human disease (206 of the isolates were from the United Kingdom). In addition, 164 of these isolates were also phage typed. The isolates were divided into two general groups: (i) unrelated isolates not known to be epidemiologically linked (n ‫؍‬ 154) and originating from food animals, foods and the environment, or humans and (ii) epidemiologically related isolates (n ‫؍‬ 53) comprised of four related groups (RGs) originating either from one farm plus the abattoir where cattle from that farm were slaughtered or from one of three different English abattoirs. PFGE was conducted with the restriction endonuclease XbaI, while for ribotyping, two restriction endonucleases (PstI and SphI) were combined to digest genomic DNAs simultaneously. The 207 E. coli O157 isolates produced 97 PFGE profiles and 51 ribotypes. The two genetic fingerprinting methods had similar powers to discriminate the 154 epidemiologically unrelated E. coli O157 isolates in the study (Simpson's index of diversity [D] ‫؍‬ 0.98 and 0.94 for PFGE typing and ribotyping, respectively). There was no correlation between the source of an isolate (healthy meat or milk animals, retail meats, or cases of human infection) and either particular PFGE or ribotype profiles or clusters. Combination of the results of both genetic fingerprinting methods produced 146 types, significantly more than when either of the two methods was used individually. Consequently, the superior discriminatory performance of the PFGE-ribotyping combination was proven in two ways: (i) by demonstrating that the majority of the E. coli O157 isolates with unrelated histories were indeed distinguishable types and (ii) by identifying some clonal groups among two of the four RGs of E. coli O157 isolates (comprising PFGE types different by just one or two bands), the relatedness of which would have remained unconfirmed otherwise.

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Comparative study of five different techniques for epidemiological typing of Escherichia coli O157

Cited by 65 publications

References 31 publications

Phage Types and Genotypes of Shiga Toxin-Producing Escherichia coli O157:H7 Isolates from Humans and Animals in Spain: Identification and Characterization of Two Predominating Phage Types (PT2 and PT8)

Phage Types and Genotypes of Shiga Toxin-Producing Escherichia coli O157:H7 Isolates from Humans and Animals in Spain: Identification and Characterization of Two Predominating Phage Types (PT2 and PT8)

Genetic relationship between Escherichia coli strains isolated from dairy mastitis and from the stable fly Stomoxys calcitrans

Combined Use of Two Genetic Fingerprinting Methods, Pulsed-Field Gel Electrophoresis and Ribotyping, for Characterization of Escherichia coli O157 Isolates from Food Animals, Retail Meats, and Cases of Human Disease

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