Detection of enterotoxigenic <i>E. coli</i> in hospitalised children with and without diarrhoea in Blantyre, Malawi

Trainor, Eamonn; Iturriza‐Gómara, Miren; Ngwira, Bagrey; Cunliffe, Nigel A.

doi:10.1179/2046905515y.0000000014

Cited by 12 publications

(15 citation statements)

References 18 publications

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“…While ETEC is the most common DEC internationally, the prevalence of ETEC in this setting was found to be considerably lower than other regions [9,18]. This result is probably due to the study inclusion criteria, as only children presenting with bloody and/or mucoid diarrhoeal illness were enrolled, whereas ETEC is most commonly associated with watery diarrhoea [7].…”

Section: Discussionmentioning

confidence: 77%

“…Diarrhoeagenic E. coli (DEC) can generally be divided into six pathotypes (enterotoxigenic E. coli, ETEC; enteroaggregative E. coli, EAEC; enteropathogenic E. coli, EPEC, enteroinvasive E. coli, EIEC; enterohemorrhagic E. coli, EHEC and shiga-toxin producing E. coli, STEC), based on speci c virulence markers that are encoded on plasmids and/or chromosomal islands [7]. ETEC, EAEC, and EPEC have all been implicated in causing diarrhoea in young children in low-middle income countries (LMICs) [8][9][10].…”

Section: Introductionmentioning

confidence: 99%

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Novel multiplex real-time PCR assays reveal a high prevalence of diarrhoeagenic Escherichia coli pathotypes in healthy and diarrhoeal children in the South of Vietnam

Duong¹,

Tuyen

et al. 2020

Preprint

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Abstract Background: Diarrhoeagenic Escherichia coli (DEC) infections are common in children in low-middle income countries (LMICs). However, detecting the various DEC pathotypes is complex as they cannot be differentiated by classical microbiology. We developed four multiplex real-time PCR assays were to detect virulence markers of six DEC pathotypes; specificity was tested using DEC controls and other enteric pathogens. PCR amplicons from the six E. coli pathotypes were purified and amplified to be used to optimize PCR reactions and to calculate reproducibility. After validation, these assays were applied to clinical samples from healthy and diarrhoeal Vietnamese children and associated with clinical data. Results: The multiplex real-time PCRs were found to be reproducible, and specific. At least one DEC variant was detected in 34.7% (978/2,815) of the faecal samples from diarrhoeal children; EAEC, EIEC and atypical EPEC were most frequent Notably, 41.2% (205/498) of samples from non-diarrhoeal children was positive with a DEC pathotype. In this population, only EIEC, which was detected in 34.3% (99/289) of diarrhoeal samples vs. 0.8% (4/498) non-diarrhoeal samples (p<0.001), was significantly associated with diarrhoea. Multiplex real-time PCR when applied to clinical samples is an efficient and high-throughput approach to DEC pathotypes. Conclusions: This approach revealed high carriage rates of DEC pathotypes among Vietnamese children. We describe a novel diagnostic approach for DEC, which provides baseline data for future surveillance studies assessing DEC burden in LMICs.

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Section: Discussionmentioning

confidence: 77%

Section: Introductionmentioning

confidence: 99%

Novel multiplex real-time PCR assays reveal a high prevalence of diarrhoeagenic Escherichia coli pathotypes in healthy and diarrhoeal children in the South of Vietnam

Duong¹,

Tuyen

et al. 2020

Preprint

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“…Further limitations of this approach are associated with issues of how pathotypes such as EPEC, EHEC, and STEC are defined. Through bacterial genomics, we know that organisms lacking either eae or stx or both may still belong to the EHEC group [9,10]. In addition, the stx genes have been found in other pathotypes of E. coli [11,12].…”

Section: Discussionmentioning

confidence: 99%

“…and shiga-toxin producing E. coli, STEC), based on specific virulence markers that are encoded on plasmids and/or chromosomal islands 7 . ETEC, EAEC, and EPEC have all been implicated in causing diarrhoea in young children in low-middle income countries (LMICs) [8][9][10] . EIEC are virtually indistinguishable from Shigella spp., which are essentially an independent genus within the broader E. coli population.…”

Section: Introductionmentioning

confidence: 99%

Novel multiplex real-time PCR assays reveal a high prevalence of diarrhoeagenic Escherichia coli pathotypes in healthy and diarrhoeal children in the South of Vietnam

Duong¹,

Tuyen

et al. 2020

Preprint

View full text Add to dashboard Cite

Abstract BackgroundDiarrhoeagenic Escherichia coli (DEC) infections are common in children in low-middle income countries (LMICs). However, detecting the various DEC pathotypes is complex as they cannot be differentiated by classical microbiology. We developed four multiplex real-time PCR assays were to detect virulence markers of six DEC pathotypes; specificity was tested using DEC controls and other enteric pathogens. PCR amplicons from the six E. coli pathotypes were purified and amplified to be used to optimize PCR reactions and to calculate reproducibility. After validation, these assays were applied to clinical samples from healthy and diarrhoeal Vietnamese children and associated with clinical data. ResultsThe multiplex real-time PCRs were found to be reproducible, and specific. At least one DEC variant was detected in 34.7% (978/2,815) of the faecal samples from diarrhoeal children; EAEC, EIEC and atypical EPEC were most frequent Notably, 41.2% (205/498) of samples from non-diarrhoeal children was positive with a DEC pathotype. In this population, only EIEC, which was detected in 34.3% (99/289) of diarrhoeal samples vs. 0.8% (4/498) non-diarrhoeal samples (p<0.001), was significantly associated with diarrhoea. Multiplex real-time PCR when applied to clinical samples is an efficient and high-throughput approach to DEC pathotypes. ConclusionsThis approach revealed high carriage rates of DEC pathotypes among Vietnamese children. We describe a novel diagnostic approach for DEC, which provides baseline data for future surveillance studies assessing DEC burden in LMICs.

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“…Many outbreaks of human gastroenteritis have been linked to the consumption of contaminated fresh vegetables and fruits (Garner and Kathariou, 2016). Enterotoxigenic E. coli, specifically serotype O157:H7, is the most common cause of diarrhea (Trainor et al, 2016), and consumption of E. coli O157:H7 contaminated cantaloupes caused an outbreak of gastroenteritis (Denis et al, 2016). Besides implementation of food safety plans with sound prerequisite programs and appropriate preventive controls for identified hazard to reduce the risk of foodborne infections, it is important to develop improved decontamination technologies for both food contact surfaces and the food itself.…”

mentioning

confidence: 99%

The Effect of Controlled-release Chlorine Dioxide on the Preservation of Grapefruit

Baldwin¹,

Ference²,

Narciso³

et al. 2017

horts

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The effect of controlled-release chlorine dioxide (ClO2) gas on the safety and quality of grapefruit was studied. The experiments were run under controlled chamber systems with inoculated fruit, and in boxed fruit under commercial conditions. For the inoculation test, fruit artificially inoculated with either Escherichia coli or Penicillium digitatum, or naturally inoculated Xanthomonas citri ssp. citri (Xcc) (fruits with citrus canker lesions), were incubated in a chamber containing a dose equivalent to 0–60 mg·L−1 of pure ClO2 as an antimicrobial agent. After 24 hours, the microbial population on treated grapefruit was significantly reduced compared with that of control fruit: a dosage of 5 mg·L−1 completely inhibit the growth of E. coli and P. digitatum, but a dosage of 60 mg·L−1 was needed to completely kill Xcc. For the simulated commercial experiment, fruit were harvested in late Oct. 2015 passed through a commercial packing line, and packed in 29 L citrus boxes. ClO2 packets were attached to the top lids with the following five treatments: fast-release, slow-release, slow/fast-release combination (each containing 14.5 mg·L−1 of pure ClO2), double dose fast-release (containing 29 mg·L−1 of ClO2), and control. After 6 weeks of storage at 10 °C (to simulate storage and transportation) + 1 week of storage at 20 °C (to simulate retail marketing), the fruit quality was evaluated. The slow-release treatment at standard dose exhibited the best antimicrobial activity, reducing total aerobic bacterial count and yeast/mold count by 0.95 and 0.94 log colony-forming units (cfu)/g of fruit, respectively, and maintained the best visual, sensory, and overall quality. However, the higher dosage treatments resulted in phytotoxicity as evidenced by peel browning.

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Detection of enterotoxigenic E. coli in hospitalised children with and without diarrhoea in Blantyre, Malawi

Abstract: The burden of ETEC infection in young Malawian children is substantial and should become a focus of diarrhoea prevention efforts in the post-rotavirus vaccine era.

Cited by 12 publications

References 18 publications

Novel multiplex real-time PCR assays reveal a high prevalence of diarrhoeagenic Escherichia coli pathotypes in healthy and diarrhoeal children in the South of Vietnam

Novel multiplex real-time PCR assays reveal a high prevalence of diarrhoeagenic Escherichia coli pathotypes in healthy and diarrhoeal children in the South of Vietnam

Novel multiplex real-time PCR assays reveal a high prevalence of diarrhoeagenic Escherichia coli pathotypes in healthy and diarrhoeal children in the South of Vietnam

The Effect of Controlled-release Chlorine Dioxide on the Preservation of Grapefruit

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