Comparison of a Shiga Toxin Enzyme-Linked Immunosorbent Assay and Two Types of PCR for Detection of Shiga Toxin-Producing
            <i>Escherichia coli</i>
            in Human Stool Specimens

Pulz, M.; Matussek, Andreas; Monazahian, Masyar; Tittel, A.; Nikolic, Elisabet; Hartmann, M.D.; Bellin, Tobias; Buer, Jan; Gunzer, Florian

doi:10.1128/jcm.41.10.4671-4675.2003

Cited by 45 publications

(46 citation statements)

References 32 publications

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“…In our study, 16 STEC strains were isolated from 8 patients. The difficulty in recovering STEC strains was previously reported by different authors (27,29,36). This may be explained by the freeze-and-thaw effect killing or inhibiting the growth of pathogens.…”

Section: Discussionmentioning

confidence: 77%

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: Discussionmentioning

confidence: 77%

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

2013

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“…A number of EIAs have been approved by the Food and Drug Administration (FDA) for the diagnosis of human STEC infections and have the ability to detect non-O157 STEC in addition to E. coli O157, in contrast to sorbi-tol-MacConkey agar (SMAC) culture, which detects only E. coli O157 (7,11). However, EIAs can have relatively high false-positive rates and have been reported to miss E. coli O157 (7,10,14). A number of nucleic acid-based methods, including multiplex, realtime PCR assays, have also been described (10,(15)(16)(17)(18)(19)(20).…”

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confidence: 99%

“…Additionally, a Luminex-based molecular assay targeting 15 stool pathogens including STEC was recently approved by the FDA (21). While these assays are more sensitive than EIAs (14,22), many of them require the use of fluorescently labeled probes, which can be prohibitively expensive for screening assays that would need to be performed thousands of times in order to accommodate the large numbers of stool specimens that are routinely submitted to many clinical microbiology laboratories.…”

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confidence: 99%

A Sensitive Multiplex, Real-Time PCR Assay for Prospective Detection of Shiga Toxin-Producing Escherichia coli from Stool Samples Reveals Similar Incidences but Variable Severities of Non-O157 and O157 Infections in Northern California

et al. 2013

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dRapid and accurate detection of Shiga toxin-producing Escherichia coli (STEC) of all serotypes from patients with diarrhea is critical for medical management and for the prevention of ongoing transmission. In this prospective study, we assessed the performance of a multiplex, real-time PCR assay targeting stx 1 and stx 2 for the detection of O157 and non-O157 STEC in diarrheal stool samples enriched in Gram-negative broth. We show that the assay is 100% sensitive (95% confidence interval [CI], 89.1% to 100%) and 98.5% specific (95% CI, 90.6% to 99.9%) based on a panel of 40 known STEC-positive specimens and 65 known negative specimens. During a 2-year postvalidation period, the assay detected more positive samples from patients in northern California than did culture and PCR testing performed at a public health reference laboratory, with a positive predictive value of 95.6% (95% CI, 87.6% to 99.1%). Serotyping data showed an incidence rate of 51.2% for non-O157 STEC strains, with 5.8% of patients (1/17) with non-O157 strains and 42.9% (6/14) with O157 strains (P ‫؍‬ 0.03) developing hemolytic-uremic syndrome. The findings from this study underscore the recommendations of the CDC for laboratories to test all diarrheal stool samples from patients with acute community-acquired diarrhea for non-O157 STEC in addition to the O157 serotype by using a sensitive assay. Additionally, a survey of 17 clinical laboratories in northern California demonstrated that nearly 50% did not screen all stool specimens for the presence of Shiga toxins, indicating that many clinical microbiology laboratories still do not routinely screen all stool specimens for the presence of Shiga toxins as recommended in the 2009 CDC guidelines. Shiga toxin-producing Escherichia coli (STEC) infections are a major cause of bacterial gastroenteritis throughout the world and can be complicated by hemorrhagic colitis and hemolyticuremic syndrome (HUS) (1, 2). Although previously the majority of reported cases were attributed to E. coli O157:H7 and many recent studies indicate higher incidence rates of HUS associated with E. coli O157:H7 (3, 4), non-O157 STEC serotypes are emerging as important etiological agents of both sporadic cases and community outbreaks of diarrhea (3-8). Most notably, in May and June 2011, a large outbreak of E. coli O104:H4-associated diarrheal illness in Germany led to HUS in over 800 patients, many of whom were adults, and ultimately resulted in 54 deaths (5, 9). This outbreak and others have underscored the fact that severe disease and HUS can occur in cases related to non-O157 or O157 E. coli strains and that adults as well as children are susceptible to these complications (8-10). Timely laboratory identification of STEC has important implications for outbreak containment and patient management, including prompt parenteral hydration, monitoring for development of HUS, and avoidance of antibiotics and antidiarrheal agents, which can exacerbate disease (1). Importantly, a number of studies have shown that STECrelated illnesses...

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“…STEC strains isolated from patients usually possess, in addition to one or more stx genes, the eae gene, encoding adhesin intimin (7,11,16,25,26,41,49). However, a subset of STEC strains associated with human disease lack eae, and among these, strains of serogroup O91 are the most common (2,7,35,37,47,48). In Germany during the last 5 years, serogroup O91 accounted for 6.4% to 11.0% of all STEC strains reported from human infections and was therefore the fourth-most-common STEC serogroup (after O157, O26, and O103) isolated (47, 48; http://www.rki.de).…”

mentioning

confidence: 99%

Shiga Toxin, Cytolethal Distending Toxin, and Hemolysin Repertoires in Clinical Escherichia coli O91 Isolates

et al. 2009

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Comparison of a Shiga Toxin Enzyme-Linked Immunosorbent Assay and Two Types of PCR for Detection of Shiga Toxin-Producing Escherichia coli in Human Stool Specimens

Cited by 45 publications

References 32 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

A Sensitive Multiplex, Real-Time PCR Assay for Prospective Detection of Shiga Toxin-Producing Escherichia coli from Stool Samples Reveals Similar Incidences but Variable Severities of Non-O157 and O157 Infections in Northern California

Shiga Toxin, Cytolethal Distending Toxin, and Hemolysin Repertoires in Clinical Escherichia coli O91 Isolates

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