Detection, Isolation, and Molecular Subtyping of
            <i>Escherichia coli</i>
            O157:H7 and
            <i>Campylobacter jejuni</i>
            Associated with a Large Waterborne Outbreak

Bopp, Dianna J.; Sauders, Brian D.; Waring, Alfred L.; Ackelsberg, Joel; Dumas, Nellie B.; Braun-Howland, Ellen B.; Dziewulski, David M.; Wallace, Barbara J.; Kelly, Molly E; Halse, Tanya A.; Musser, Kimberlee A.; Smith, Perry F.; Morse, Dale L.; Limberger, Ronald J.

doi:10.1128/jcm.41.1.174-180.2003

Cited by 110 publications

(68 citation statements)

References 55 publications

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“…This finding was previously described by many studies (37)(38)(39)(40). Among them, Hedican et al performed a sentinel surveillance of STEC infections in Minnesota and found 22 coinfected patients among 302 STEC-positive patients (40).…”

Section: Discussionsupporting

confidence: 59%

Comparison of Three Different Methods for Detection of Shiga Toxin-Producing Escherichia coli in a Tertiary Pediatric Care Center

2013

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Shiga toxin-producing Escherichia coli (STEC) is a well-known cause of sporadic and epidemic food-borne gastroenteritis. A low infectious dose, approximately 10 microorganisms, is sufficient to cause disease that may lead to hemolytic-uremic syndrome. The objective of this study was to compare the performances of an in-house real-time PCR, a commercial enzyme immunoassay (EIA) (Premier EHEC; Meridian Bioscience), and culture on sorbitol MacConkey agar for the detection of STEC in a tertiary care pediatric hospital. Of 632 stool samples tested, 21 were positive for STEC. All were detected by PCR, 6 were detected by EIA, and only 5 O157 STEC isolates were identified by culture. Among the 15 specimens falsely negative by EIA, there were 9 Stx1, 2 Stx2, and 4 Stx1 and Stx2 STEC isolates. The latter group included 2 O157 STEC isolates that would have been missed if only EIA had been performed. To our knowledge, this is the first prospective study performed in a pediatric hospital which demonstrates the superiority of PCR over EIA for the detection of STEC. We conclude that PCR is specific and more sensitive than EIA. PCR should be considered for routine use in clinical settings where molecular detection facilities are available. Its lower limit of detection, equivalent to the infectious dose, is an obvious advantage for patient care and public health surveillance.

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Section: Discussionsupporting

confidence: 59%

Comparison of Three Different Methods for Detection of Shiga Toxin-Producing Escherichia coli in a Tertiary Pediatric Care Center

2013

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“…C. jejuni is highly prevalent in poultry (4,5,19,31), and poultry products are often assumed to be the source of most C. jejuni infections (22,30,49,58,62), although poultry isolates may not all be equally pathogenic (25,51). Other sources, such as untreated water (7,15,29) and unpasteurized milk (18,34), can also lead to campylobacteriosis. C. jejuni infections are mainly sporadic, although occasional outbreaks can occur (23,54).…”

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confidence: 99%

Extended Multilocus Sequence Typing System for Campylobacter coli , C. lari , C. upsaliensis , and C. helveticus

On²,

et al. 2005

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A multilocus sequence typing (MLST) system has been reported previously for Campylobacter jejuni to both differentiate strains and identify clonal lineages. However, sequence variation at the MLST loci prevents its use for closely related Campylobacter species. We describe herein an expanded MLST method to include three clinically relevant Campylobacter species, C. coli, C. lari, and C. upsaliensis, and a fourth Campylobacter species, C. helveticus. The C. coli and C. helveticus methods use the same seven C. jejuni loci (aspA, atpA, glnA, gltA, glyA, pgm, and tkt); however, adk and pgi were substituted for aspA and gltA in C. lari and for gltA and pgm in C. upsaliensis. Multiple C. coli (n ‫؍‬ 57), C. lari (n ‫؍‬ 20), C. upsaliensis (n ‫؍‬ 78), and C. helveticus (n ‫؍‬ 9) isolates, representing both clinical and environmental sources, were typed. All four species were genetically diverse: the majority (>80%) of the isolates had unique sequence types (STs). Using this method, mixed C. lari, C. upsaliensis, and C. helveticus isolates were identified; upon separation, each isolate was shown to contain two strains of the same species with distinct STs. Additionally, the expanded MLST method was able to detect potential lateral transfer events between C. jejuni and either C. coli or C. lari and between C. upsaliensis and C. helveticus. Thus, the expanded MLST method will prove useful in differentiating strains of five Campylobacter species, identifying mixed Campylobacter cultures, and detecting genetic exchange within the genus.

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“…Pulsed-field gel electrophoresis (PFGE) is the most common molecular biologybased method used for the subtyping of STEC strains. This method is considered the "gold standard" for the subtyping of STEC strains, due to its high power of discrimination and reproducibility, and has been successfully used all over the world (10,11,16,24). Moreover, the standardization of PFGE analysis for epidemiological purposes has been achieved in the United States to facilitate the subtyping of bacterial food-borne pathogens among different laboratories (39).…”

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confidence: 99%

Genetic Heterogeneity of Shiga Toxin-Producing Escherichia coli Strains Isolated in São Paulo, Brazil, from 1976 through 2003, as Revealed by Pulsed-Field Gel Electrophoresis

et al. 2006

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The pulsed-field gel electrophoresis (PFGE) patterns of 46 Shiga toxin-producing Escherichia coli (STEC) strains isolated in São Paulo, Brazil, during the period from 1976 to 2003 were compared with those found among 30 non-STEC strains that carried eae and that belonged to the same serogroups as the STEC strains. All except two of the STEC and non-STEC strains of human origin were from sporadic and unrelated cases of infection; two O111 strains originated from the same patient. Multiple PFGE patterns were found among STEC strains of distinct serotypes. Moreover, the PFGE restriction patterns of STEC strains differed substantially from those observed among non-STEC strains of the same serogroup except serotype O26 strains. Based on the indistinguishable PFGE pattern for two O157:H7 STEC strains isolated in the same geographic area at an interval of approximately 15 days and toxin profile data, the first occurrence of an O157:H7 outbreak in Brazil during that period can be suggested. In general, a close relationship between types of intimin, serotypes, and diarrheagenic groups of E. coli was observed. This is the first time that a large collection of STEC strains from Brazil has been analyzed, and a great genetic diversity was shown among O157:H7 and non-O157:H7 STEC strains isolated in São Paulo, Brazil.

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Detection, Isolation, and Molecular Subtyping of Escherichia coli O157:H7 and Campylobacter jejuni Associated with a Large Waterborne Outbreak

Cited by 110 publications

References 55 publications

Comparison of Three Different Methods for Detection of Shiga Toxin-Producing Escherichia coli in a Tertiary Pediatric Care Center

Comparison of Three Different Methods for Detection of Shiga Toxin-Producing Escherichia coli in a Tertiary Pediatric Care Center

Extended Multilocus Sequence Typing System for Campylobacter coli , C. lari , C. upsaliensis , and C. helveticus

Genetic Heterogeneity of Shiga Toxin-Producing Escherichia coli Strains Isolated in São Paulo, Brazil, from 1976 through 2003, as Revealed by Pulsed-Field Gel Electrophoresis

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