BMC Microbiology | |
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 | |
Ellen Higginson1  Stephen Baker1  Hoang Le Phuc2  Nguyen Minh Ngoc3  Iruka Okeke4  Claire Jenkins5  Ha Thanh Tuyen6  Vu Thuy Duong6  James I. Campbell6  Le Thi Quynh Nhi6  Pham Van Minh6  Le Thi Phuong Tu6  Tran Thi Hong Chau6  Lu Lan Vi7  | |
[1] Cambridge Institute of Therapeutic Immunology & Infectious Disease (CITIID) Department of Medicine, Cambridge Biomedical Campus, University of Cambridge;Children’s Hospital 1;Children’s Hospital 2;Department of Pharmaceutical Microbiology, Faculty of Pharmacy, University of Ibadan;National Infection Service, Public Health England;The Hospital for Tropical Diseases, Wellcome Trust Major Overseas Programme, Oxford University Clinical Research Unit;The Hospital for Tropical Diseases; | |
关键词: ETEC; EAEC; EIEC; EPEC; EHEC; Multiplex real-time PCR; | |
DOI : 10.1186/s12866-020-01878-5 | |
来源: DOAJ |
【 摘 要 】
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 < 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.
【 授权许可】
Unknown