Evaluation of Pulsed-field Gel Electrophoresis (PFGE) for Yersinia enterocolitica Molecular Epidemiology Investigations

2004 ◽  
pp. 375-378 ◽  
Author(s):  
Marina Marranzano ◽  
Maria A. Coniglio ◽  
Luisa Mauro
Keyword(s):  
Molecular Epidemiology ◽  
Yersinia Enterocolitica ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field

scholarly journals Identification and Characterization of Pathogenic Yersinia enterocolitica Isolates by PCR and Pulsed-Field Gel Electrophoresis

2005 ◽  
Vol 71 (7) ◽  
pp. 3674-3681 ◽  
Author(s):  
S. Thisted Lambertz ◽  
M.-L. Danielsson-Tham
Keyword(s):  
Multiplex Pcr ◽  
Yersinia Enterocolitica ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Virulence Plasmid ◽  
Pork Meat ◽  
Pulsed Field ◽  
Pork Products ◽  
Food Borne ◽  
Pork Samples

ABSTRACT Approximately 550 to 600 yersiniosis patients are reported annually in Sweden. Although pigs are thought to be the main reservoir of food-borne pathogenic Yersinia enterocolitica, the role of pork meat as a vehicle for transmission to humans is still unclear. Pork meat collected from refrigerators and local shops frequented by yersiniosis patients (n = 48) were examined for the presence of pathogenic Yersinia spp. A combined culture and PCR method was used for detection, and a multiplex PCR was developed and evaluated as a tool for efficient identification of pathogenic food and patient isolates. The results obtained with the multiplex PCR were compared to phenotypic test results and confirmed by pulsed-field gel electrophoresis (PFGE). In all, 118 pork products (91 raw and 27 ready-to-eat) were collected. Pathogenic Yersinia spp. were detected by PCR in 10% (9 of 91) of the raw pork samples (loin of pork, fillet of pork, pork chop, ham, and minced meat) but in none of the ready-to-eat products. Isolates of Y. enterocolitica bioserotype 4/O:3 were recovered from six of the PCR-positive raw pork samples; all harbored the virulence plasmid. All isolates were recovered from food collected in shops and, thus, none were from the patients' home. When subjected to PFGE, the six isolates displayed four different NotI profiles. The same four NotI profiles were also present among isolates recovered from the yersiniosis patients. The application of a multiplex PCR was shown to be an efficient tool for identification of pathogenic Y. enterocolitica isolates in naturally contaminated raw pork.

scholarly journals Evaluation of pulsed-field gel electrophoresis of genomic restriction fragments in the discrimination of Yersinia enterocolitica O[ratio ]3

1998 ◽  
Vol 121 (3) ◽  
pp. 579-586 ◽  
Author(s):  
K. ASPLUND ◽  
T. JOHANSSON ◽  
A. SIITONEN
Keyword(s):  
Yersinia Enterocolitica ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Restriction Fragments ◽  
Pulsed Field ◽  
Different Types

One hundred and six Yersinia enterocolitica serogroup O[ratio ]3, biotype 4 isolated from human and porcine samples in 1984 and in the years 1993–5 were examined by pulsed-field gel electrophoresis (PFGE). The genomic profiles produced by the enzymes NotI and XbaI were studied. Sixteen (A–P) and 8 (1·8) different pulsotypes were obtained, respectively. By combining the pulsotypes produced by both NotI and XbaI 24 different types were distinguished. The two major types, designated as A1 and B1, comprised 36% of all strains tested. The proportions of pulsotypes A1 and B1 were, 35·9 and 25·6%, respectively, among strains isolated in 1984. The corresponding figures among the strains isolated in 1993–5 were 35·8 and 41·8%. Nine pulsotypes were found only in 1984 and nine only in 1993–5. The proportions of the major pulsotypes, A1 and B1, in human isolates were 42·9 and 35·7% and in porcine isolates 22·2 and 36·1% respectively. Six types were found among both human and porcine isolates, 8 only among human strains and 10 only among porcine strains.

scholarly journals Molecular epidemiology of Salmonella enteritidis phage type 1b and 6a isolates in Portugal

2003 ◽  
Vol 131 (1) ◽  
pp. 607-612 ◽  
Author(s):  
A. R. SOARES ◽  
J. MACHADO
Keyword(s):  
Molecular Epidemiology ◽  
Gel Electrophoresis ◽  
Salmonella Enterica ◽  
Genomic Dna ◽  
Salmonella Enteritidis ◽  
Phage Type ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Main Pattern

Salmonella enterica serotype Enteritidis is an important serovar comprising 76% of salmonella isolates in Portugal in 2001.For better understand the epidemiology of salmonellosis, a total of 47 isolates of S. Enteritidis phage type (PT) 1b and 6a were analysed by pulsed-field gel electrophoresis (PFGE) and genomic DNA was subjected to macro restriction with XbaI. For PT1b isolates, only three different patterns were observed, and PT6a showed a total of 10 digestion patterns. Curiously, the main pattern among PT1b isolates seams quite similar to main pattern of PT6a isolates, but when the two patterns were analysed with Bionumerics, we observed that they exhibited some differences. It was concluded that, in 2001, there was one predominant pattern for PT1b and PT6a and, possibly, we were in presence of clonal strains that exists all over the country.

scholarly journals Molecular Epidemiology of Penicillin-Resistant and Ciprofloxacin-Resistant Streptococcus pneumoniae in Canada

2003 ◽  
Vol 47 (2) ◽  
pp. 804-808 ◽  
Author(s):  
Kimberly A. Nichol ◽  
George G. Zhanel ◽  
Daryl J. Hoban
Keyword(s):  
Streptococcus Pneumoniae ◽  
Molecular Epidemiology ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Penicillin Resistance ◽  
Genotypic Variability ◽  
Pulsed Field ◽  
Ciprofloxacin Resistance

ABSTRACT Eighty-nine penicillin- and ciprofloxacin-resistant Streptococcus pneumoniae isolates were evaluated by serotyping and pulsed-field gel electrophoresis. Although penicillin-resistant isolates demonstrated considerable homogeneity, resistance to ciprofloxacin did not correlate with a reduction in genotypic variability. These results suggest that, unlike that of penicillin resistance, the spread of S. pneumoniae ciprofloxacin resistance in Canada is currently not attributable to clonal dissemination.

scholarly journals Use of Pulsed-Field Gel Electrophoresis for Molecular Epidemiologic and Population Genetic Studies ofMycobacterium tuberculosis †

1999 ◽  
Vol 37 (6) ◽  
pp. 1927-1931 ◽  
Author(s):  
Samir P. Singh ◽  
Hugh Salamon ◽  
Carol J. Lahti ◽  
Mehran Farid-Moyer ◽  
Peter M. Small
Keyword(s):  
Genetic Diversity ◽  
Molecular Epidemiology ◽  
Gel Electrophoresis ◽  
Population Biology ◽  
Rflp Analysis ◽  
Analytical Techniques ◽  
Restriction Enzymes ◽  
Pulsed Field Gel Electrophoresis ◽  
Molecular Biology Technique ◽  
Pulsed Field

Pulsed-field gel electrophoresis (PFGE) is a powerful molecular biology technique which has provided important insights into the epidemiology and population biology of many pathogens. However, few studies have used PFGE for the molecular epidemiology ofMycobacterium tuberculosis. A laboratory protocol was developed to determine the typeability, stability, and reproducibility of PFGE typing of M. tuberculosis. Formal data-analytical techniques were used to assess the genetic diversity elucidated by PFGE analyses using four separate restriction enzymes and by IS6110 RFLP analyses, as well as to assess the concordance among these typing methods. One hundred epidemiologically characterized clinical isolates of M. tuberculosis were genotyped with four different PFGE enzymes (AseI, DraI,SpeI, and XbaI), as well as by RFLP analysis with IS6110. Identical patterns were found among 34 isolates known to be genetically related, suggesting that the PFGE protocol is robust and reproducible. Among 66 isolates representing population-sampled cases, heterozygosity and information content dependency estimates indicate that all five genotyping systems capture quantitatively similar levels of genetic diversity. Nevertheless, comparisons between PFGE analyses and IS6110 typing reveals that PFGE provided more discrimination among isolates with fewer than five copies of IS6110 and less clustering in isolates with five or more copies. The comparisons confirm the hypothesis that the resolution of IS6110 RFLP genotyping is dependent upon the number of IS6110 elements in the genome of isolates. The general concordance among the results obtained with four independent enzymes suggests that M. tuberculosis is a clonal organism. The availability of a robust genotyping technique largely independent of repetitive elements has implications for the molecular epidemiology of M. tuberculosis.

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