1. 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
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Vu Thuy Duong, Le Thi Phuong Tu, Ha Thanh Tuyen, Le Thi Quynh Nhi, James I. Campbell, Pham Van Minh, Hoang Le Phuc, Tran Thi Hong Chau, Nguyen Minh Ngoc, Lu Lan Vi, Claire Jenkins, Iruka Okeke, Ellen Higginson, and Stephen Baker more...
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ETEC ,EAEC ,EIEC ,EPEC ,EHEC ,Multiplex real-time PCR ,Microbiology ,QR1-502 - Abstract
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% (978/2815) 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 more...
- Published
- 2020
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