[Show abstract][Hide abstract] ABSTRACT: Phage typing and DNA macrorestriction fragment analysis by pulsed-field gel electrophoresis (PFGE) were evaluated for use in the epidemiological subtyping of Escherichia coli serogroup O157 strains isolated in Ontario, Canada. Among 30 strains isolated from patients with sporadic cases of infection, 22 distinct XbaI macrorestriction patterns were identified and 17 strains exhibited unique PFGE patterns. In contrast, phage typing identified only seven different phage types and 17 strains belonged to the same phage type. A total of 25 phage type-macrorestriction pattern combinations were identified among the strains from patients with sporadic cases of infection. PFGE subtyping differentiated between unrelated strains that exhibited the same phage type, and in one group of strains, phage typing differentiated between strains of the same PFGE subtype. Both typing procedures correctly identified outbreak-related isolates as belonging to the same type in four separate outbreaks. Each outbreak strain was characterized by a distinct macrorestriction pattern, while phage typing subdivided the outbreak strains into only three different types. A small percentage of outbreak-related isolates had PFGE patterns that differed slightly (one or two DNA fragment differences) from that of the outbreak strain. On the other hand, each isolate from the same outbreak belonged to the same phage type as that of the outbreak strain. We conclude that phage typing and PFGE fingerprinting represent complementary procedures for the subtyping of E. coli serogroup O157 and that the combined use of these procedures provides optimal discrimination.
Journal of Clinical Microbiology 07/2000; 38(6):2366-8. · 3.99 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The utility of phage typing, pulsed-field gel electrophoresis (PFGE), and plasmid profile analysis was compared, to differentiate between Canadian Escherichia coli O157:H7 strains of human (n = 27) and cattle (n = 24) origin. The diversity indices for phage typing, plasmid analysis and PFGE were 0.85, 0.69 and 0.93, respectively. PFGE and phage typing were also applied to study the role of direct transmission of E. coli O157:H7 from cattle to humans on isolates collected from two separate farm outbreaks. PFGE showed that more than one E. coli O157:H7 strain with varying PFGE DNA subtype profiles, may be responsible for an outbreak, and that more than one E. coli O157:H7 subtype may be circulating on a particular farm at any one time. To our knowledge, this is one of the first reports where PFGE typing was used to verify the direct transmission of E. coli O157:H7 from cattle to humans.
Epidemiology and Infection 09/1999; 123(1):17-24. DOI:10.1017/S0950268899002551 · 2.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In September 1994, a complaint was registered at a public health unit concerning a cheese product. In addition, public health laboratories in Ontario reported an increase in the number of isolates of Salmonella berta from patients with diarrhoeal illness. A clinical, environmental and laboratory investigation was initiated to determine the nature of this outbreak. Isolates of Salmonella berta were compared using large fragment genomic fingerprinting by pulsed-field gel electrophoresis (PFGE). By late October, 82 clinical cases had been identified including 35 confirmed, 44 suspected and 3 secondary. The investigation linked illness to consumption of an unpasteurized soft cheese product produced on a farm and sold at farmers' markets. Subtyping results of patient, cheese and chicken isolates were indistinguishable, suggesting that the cheese was contaminated by chicken carcasses during production. The outbreak illustrates the potential role of uninspected home-based food producers and of cross-contamination in the transmission of foodborne bacterial pathogens.
Epidemiology and Infection 02/1998; 120(1):29-35. DOI:10.1017/S0950268897008376 · 2.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Recent policies of early discharge of postpartum mothers and their infants has raised concerns of possible decreased sensitivity in Guthrie bacterial inhibition assay (BIA) phenylketonuria (PKU) screening resulting in missed cases. In order to assess the potential impact of early discharge from hospital on neonatal screening for PKU and its variants, we performed 18 standard BIA screening tests on 11 newborn infants with the disease. Blood spot samples were collected from 1 to 24 h after birth and were analyzed at the Ontario Ministry of Health newborn screening laboratory according to the routine screening protocol. Except for one 4-hour postnatal sample from an infant with 'non-PKU mild hyperphenylalaninemia' (MHP) all blood samples showed phenylalanine levels > or = 240 mumol/l, irrespective of the age of the baby. During our 29 year experience with neonatal PKU screening (3.9 million infants tested), employing a cutoff blood phenylalanine of 240 mumol/l in blood spots obtained at > or = 24 h of age, only two biological false negative (one confirmed) tests were discovered in infants subsequently shown to have classical PKU: another three false negative tests were discovered in sibs of infants with MHP. The sensitivity of the screening test was 99.2% for infants with classical and mild PKU. Ascertainment of patients with MHP is unknown and is very likely incomplete. Over a 3-year period (1992-4) the specificity of the test was 99.9% for those screened after 24 h. The positive predictive value was 12.8%. Although early discharge may have an impact on other screened diseases, we conclude, from our studies, that early discharge may not affect the detection of infants with classical and mild (atypical) PKU, but would probably increase the number of infants with MHP missed using the BIA and a cutoff level of 240 mumol/l. Because of our experience and that of others, we recommend that neonates be at least 12 h of age before initial BIA PKU screening be carried out. To confirm this recommendation further prospective studies should be initiated.
Early Human Development 01/1997; 47(1):87-96. DOI:10.1016/S0378-3782(96)01846-4 · 1.79 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Enterohemorrhagic Escherichia coli (EHEC) strains detected with DNA probes (for virulence plasmid and Shiga-like toxins) from subjects with hemolytic-uremic syndrome (n = 19) or diarrhea (n = 41) or asymptomatic carriers (n = 29) were examined for sorbitol fermentability, as were enterotoxigenic (n = 40), enteropathogenic (n = 40), and enteroinvasive (n = 40) E. coli and urinary tract infection (n = 40) strains and normal flora E. coli strains (n = 40). Sorbitol negativity was common only in EHEC, particularly among strains from severe clinical infections. All 19 EHEC strains from patients with hemolytic-uremic syndrome, irrespective of O:H serotype or Shiga-like toxin genotype, were sorbitol negative.
Journal of Clinical Microbiology 09/1995; 33(8):2199-201. · 3.99 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Yersinia enterocolitica has emerged as an enteropathogen associated with several types of human infections that often require antimicrobial therapy, but little is known about the antimicrobial susceptibilities of pathogenic strains isolated from humans in Canada. To determine the present patterns of antimicrobial susceptibility, to identify changes in these patterns that occurred during the past two decades, and to investigate the relationships between O serotypes and patterns of susceptibility, we tested a total of 1,105 pathogenic Y. enterocolitica strains isolated during 1972 to 1976, 1980, 1985, and 1990 for their susceptibilities to 22 antimicrobial agents. Susceptibility testing was performed by using a single breakpoint concentration in agar procedure. The results showed that all strains were susceptible to ciprofloxacin and piperacillin, and 98% or more of the strains from each period were susceptible to trimethoprim, sulfamethoxazole, cotrimoxazole, tetracycline, chloramphenicol, cefamandole, cefotaxime, aztreonam, and four aminoglycosides. In contrast, all strains were nonsusceptible to erythromycin, furazolidone, and clindamycin and 90% or more of the strains from each period were nonsusceptible to ampicillin, carbenicillin, ticarcillin, and cephalothin. Strains belonging to serotypes O:3, O:5,27, and O:8 had different patterns of susceptibility to ampicillin, carbenicillin, ticarcillin, and amoxicillin-clavulanic acid. No major difference in susceptibility was observed between any of the groups of human or animal strains included in the study, but nonsusceptibility to tetracycline increased from 0.4% in 1985 to 2% in 1990 in human strains isolated in those years. Our results indicate that between 1972 and 1990 there was no marked decrease in susceptibility to agents commonly used for therapy among pathogenic Y. enterocolitica strains isolated in Canada.
[Show abstract][Hide abstract] ABSTRACT: The distribution of the Escherichia coli attaching and effacing (eae) gene in strains of verotoxin-producing E. coli (VTEC) isolated from cattle and humans was studied. The majority of strains isolated from humans with bloody diarrhoea or HUS and cattle with severe diarrhoea were eae positive (82 and 83% respectively). In contrast, 59% of VTEC isolated from asymptomatic cattle were eae negative and of the remaining 41% that were eae positive, the majority were serotype O157. H7. The nucleotide sequence of the 3' end of the eae gene of enteropathogenic E. coli (EPEC) of serotype O55. H7 was found to be almost identical to that of serotype O157. H7. Specific primers are described which detect the eae sequences of VTEC serotypes O157. H7, O157. H-, and EPEC serotypes O55. H7 and O55. H-. The nucleotide sequence of the 3' end of the eae gene of serotype O111. H8 differed significantly from that of O157. H7. Primers were developed to specifically identify the eae sequences of VTEC serotypes O111. H- and O111. H8. We conclude that whereas the majority of VTEC associated with disease in cattle and humans possess the eae gene, the gene itself may not be necessary to produce haemorrhagic colitis and HUS. Sequence heterogeneity in the 3' end of eae alleles of VTEC permits specific identification of subsets of these organisms.
Epidemiology and Infection 07/1994; 112(3):449-61. DOI:10.1017/S0950268800051153 · 2.54 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Restriction fragment length polymorphisms (RFLPs) of genomic DNAs from 49 clinical isolates of Shigella sonnei were analyzed by using a modified restriction endonuclease analysis procedure to investigate the genetic variability of this species. After cleavage with the restriction enzyme HaeIII or RsaI, DNA samples were electrophoresed in polyacrylamide gels and the RFLP patterns were visualized by silver staining. The results showed that among 20 strains associated with sporadic cases of infection in three Canadian provinces, 15 distinct RFLP patterns were revealed by HaeIII digestion and 12 distinct patterns were revealed by RsaI digestion. In contrast, the RFLP patterns of individual isolates within six groups of epidemiologically related isolates were identical to each other but distinct from those of unrelated isolates, and these patterns could be used to determine the genetic relationships between isolates associated with separate outbreaks of shigellosis. Our results indicate that the modified restriction endonuclease analysis technique represents a rapid, reproducible, and highly discriminatory method for the molecular typing of this species.
Journal of Clinical Microbiology 07/1994; 32(6):1427-30. · 3.99 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A single clone, Neisseria meningitidis serogroup C (C:2a:P1.2), was isolated from seven patients during a cluster of cases of meningococcal disease in Ontario in 1989. To determine whether the clone was present in asymptomatic individuals in the same population, pharyngeal swabs were taken from 7% (644 of 9125) of residents who were vaccinated during the outbreak. Rates of isolation of Neisseria species were also compared to those in two other geographical areas which did not have an elevated incidence of meningococcal disease. The rate of carriage of N meningitidis in the asymptomatic individuals sampled was between 1.9% and 5.4%. The clone isolated from patients was not present among the carrier strains as determined by sero- and subtyping and electrophoretic analysis of metabolic enzymes. Age greater than six years was the only factor associated with colonization with N meningitidis.
The Canadian journal of infectious diseases = Journal canadien des maladies infectieuses 01/1992; 3(2):60-4. · 0.76 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Multilocus enzyme electrophoresis was used to characterize 378 isolates of Neisseria meningitidis serogroup C recovered during a period of an increase in group C meningococcal disease in Canada. Thirty-four enzyme electrophoretic types were found among the isolates, which were predominantly (96.0%) serotype 2a. One clone (ET 15), characterized by a rarely occurring allele for the enzyme fumarase, was responsible for a focal outbreak in Ontario followed by the spread of group C disease across the province. This clone, which occurred infrequently among strains isolated in 1986, accounted for over 65% of group C strains associated with meningococcal disease in Canada in 1990.
Journal of Clinical Microbiology 12/1991; 29(11):2489-93. · 3.99 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A large outbreak of Campylobacter jujuni gastroenteritis attributed to contamination of an unchlorinated municipal water system was investigated. Unlike most previous summer outbreaks, this one began in early spring and was attributed to meltwater entering one or more municipal wells. 241 suspected cases were documented, but retrospective information from local health care workers suggested a much larger outbreak. 45 laboratory-confirmed cases participated in a case-control study which showed a significant association between infection and amount of town water consumed. Stool specimens from 29 patients were studied with detailed serotyping by the method of Lior, with eight known serotypes and one previously unknown one identified. It is concluded that intensive surveillance of water quality during periods of spring runoff is essential, and that timely reporting of disease outbreak patterns in emergency department settings is necessary to protect the public's health.
Canadian journal of public health. Revue canadienne de santé publique 01/1991; 82(1):27-31. · 1.02 Impact Factor