Rapid O serogroup identification of the ten most clinically relevant STECs by Luminex microbead-based suspension array

Lin, Andrew; Nguyen, Lam T.; Lee, Teresa; Clotilde, Laurie M.; Kase, Julie A.; Son, Injoon; Carter, John M.; Lauzon, Carol R.

doi:10.1016/j.mimet.2011.07.019

Cited by 57 publications

(37 citation statements)

References 27 publications

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“…Serotype information for all 435 E. coli isolates used in this study was provided by the contributors of the strains. However, serogroups O26, O45, O91, O103, O104, O111, O113, O121, O128, O145, and O157 were confirmed via a Luminex microbead-based suspension array with the Bio-Plex 200 instrument (Luminex Corporation, Austin, TX) as described previously (57). Antisera from Statens Serum Institut (MiraVista, Indianapolis, IN) were used to confirm the BioPlex results (58) for non-O157 E. coli (except serogroup O104), and the E. coli O157:H7 latex test kit (Remel, Lenexa, KS) was used to confirm O157.…”

Section: Methodsmentioning

confidence: 99%

Prevalence of Hemolysin Genes and Comparison of ehxA Subtype Patterns in Shiga Toxin-Producing Escherichia coli (STEC) and Non-STEC Strains from Clinical, Food, and Animal Sources

Lorenz

Son

Maounounen-Laasri

et al. 2013

Appl Environ Microbiol

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c Shiga toxin-producing Escherichia coli (STEC) belonging to certain serogroups (e.g., O157 and O26) can cause serious conditions like hemolytic-uremic syndrome (HUS), but other strains might be equally pathogenic. While virulence factors, like stx and eae, have been well studied, little is known about the prevalence of the E. coli hemolysin genes (hlyA, ehxA, e-hlyA, and sheA) in association with these factors. Hemolysins are potential virulence factors, and ehxA and hlyA have been associated with human illness, but the significance of sheA is unknown. Hence, 435 E. coli strains belonging to 62 different O serogroups were characterized to investigate gene presence and phenotypic expression of hemolysis. We further investigated ehxA subtype patterns in E. coli isolates from clinical, animal, and food sources. While sheA and ehxA were widely distributed, e-hlyA and hlyA were rarely found. Most strains (86.7%) were hemolytic, and significantly more hemolytic (95%) than nonhemolytic strains (49%) carried stx and/or eae (P < 0.0001). ehxA subtyping, as performed by using PCR in combination with restriction fragment length polymorphism analysis, resulted in six closely related subtypes (>94.2%), with subtypes A/D being eae-negative STECs and subtypes B, C, E, and F eae positive. Unexpectedly, ehxA subtype patterns differed significantly between isolates collected from different sources (P < 0.0001), suggesting that simple linear models of exposure and transmission need modification; animal isolates carried mostly subtypes A/C (39.3%/42.9%), food isolates carried mainly subtype A (81.9%), and clinical isolates carried mainly subtype C (66.4%). Certain O serogroups correlated with particular ehxA subtypes: subtype A with O104, O113, and O8; B exclusively with O157; C with O26, O111, and O121.

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Section: Methodsmentioning

confidence: 99%

Prevalence of Hemolysin Genes and Comparison of ehxA Subtype Patterns in Shiga Toxin-Producing Escherichia coli (STEC) and Non-STEC Strains from Clinical, Food, and Animal Sources

Lorenz

Son

Maounounen-Laasri

et al. 2013

Appl Environ Microbiol

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“…With the extensive sequencing of the O-antigen-coding locus of various E. coli strains, serogroup-specific oligonucleotide pairs can be used in PCR assays (reviewed in reference 10). Serotyping methods have been developed to detect STEC using microarrays, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and microbeads (372)(373)(374). Quantitative PCR (qPCR) panels have also been developed to look at multiple genes, such as rfbE (O157 antigen), stx, eae, ehx, fliC, and O-antigen genes to profile for certain STEC isolates (375).…”

Section: Clinical Considerationsmentioning

confidence: 99%

Recent Advances in Understanding Enteric Pathogenic Escherichia coli

et al. 2013

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SUMMARY Although Escherichia coli can be an innocuous resident of the gastrointestinal tract, it also has the pathogenic capacity to cause significant diarrheal and extraintestinal diseases. Pathogenic variants of E. coli (pathovars or pathotypes) cause much morbidity and mortality worldwide. Consequently, pathogenic E. coli is widely studied in humans, animals, food, and the environment. While there are many common features that these pathotypes employ to colonize the intestinal mucosa and cause disease, the course, onset, and complications vary significantly. Outbreaks are common in developed and developing countries, and they sometimes have fatal consequences. Many of these pathotypes are a major public health concern as they have low infectious doses and are transmitted through ubiquitous mediums, including food and water. The seriousness of pathogenic E. coli is exemplified by dedicated national and international surveillance programs that monitor and track outbreaks; unfortunately, this surveillance is often lacking in developing countries. While not all pathotypes carry the same public health profile, they all carry an enormous potential to cause disease and continue to present challenges to human health. This comprehensive review highlights recent advances in our understanding of the intestinal pathotypes of E. coli .

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“…However, the majority of studies have found asymmetrical PCR to improve signal strength (Lin et al, 2011, Deregt et al, 2006b, Das et al, 2006, Spiro et al, 2000. We also found that asymmetric PCR, using a higher ratio of labelled primer to unlabelled primer, improved the strength of the probe signal.…”

Section: Protocol Optimisationsupporting

confidence: 52%

Development of multiplex suspension microarrays for the detection of alphaviruses and flaviviruses of public health significance

Fischer¹

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Rapid O serogroup identification of the ten most clinically relevant STECs by Luminex microbead-based suspension array

Cited by 57 publications

References 27 publications

Prevalence of Hemolysin Genes and Comparison of ehxA Subtype Patterns in Shiga Toxin-Producing Escherichia coli (STEC) and Non-STEC Strains from Clinical, Food, and Animal Sources

Prevalence of Hemolysin Genes and Comparison of ehxA Subtype Patterns in Shiga Toxin-Producing Escherichia coli (STEC) and Non-STEC Strains from Clinical, Food, and Animal Sources

Recent Advances in Understanding Enteric Pathogenic Escherichia coli

Development of multiplex suspension microarrays for the detection of alphaviruses and flaviviruses of public health significance

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