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
<p><strong>Background:</strong> Diarrhoeagenic <em>Escherichia coli</em> (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...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
BioMed Central
2020
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| _version_ | 1797086319249195008 |
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| author | Duong, VT Tu, LTP Tuyen, HT Nhi, LTQ Campbell, JI Van Minh, P Le Phuc, H Chau, TTH Ngoc, NM Vi, LL Jenkins, C Okeke, I Higginson, E Baker, S |
| author_facet | Duong, VT Tu, LTP Tuyen, HT Nhi, LTQ Campbell, JI Van Minh, P Le Phuc, H Chau, TTH Ngoc, NM Vi, LL Jenkins, C Okeke, I Higginson, E Baker, S |
| author_sort | Duong, VT |
| collection | OXFORD |
| description | <p><strong>Background:</strong> Diarrhoeagenic <em>Escherichia coli</em> (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 <em>E. coli</em> 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.</p>
<p><strong>Results:</strong> 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 (<em>p</em> < 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.</p>
<p><strong>Conclusions:</strong> 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.</p> |
| first_indexed | 2024-03-07T02:20:18Z |
| format | Journal article |
| id | oxford-uuid:a3aff6bd-50ed-457a-91c6-d86617b5b51a |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:20:18Z |
| publishDate | 2020 |
| publisher | BioMed Central |
| record_format | dspace |
| spelling | oxford-uuid:a3aff6bd-50ed-457a-91c6-d86617b5b51a2022-03-27T02:28:53ZNovel multiplex real-time PCR assays reveal a high prevalence of diarrhoeagenic Escherichia coli pathotypes in healthy and diarrhoeal children in the south of VietnamJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:a3aff6bd-50ed-457a-91c6-d86617b5b51aEnglishSymplectic ElementsBioMed Central2020Duong, VTTu, LTPTuyen, HTNhi, LTQCampbell, JIVan Minh, PLe Phuc, HChau, TTHNgoc, NMVi, LLJenkins, COkeke, IHigginson, EBaker, S<p><strong>Background:</strong> Diarrhoeagenic <em>Escherichia coli</em> (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 <em>E. coli</em> 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.</p> <p><strong>Results:</strong> 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 (<em>p</em> < 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.</p> <p><strong>Conclusions:</strong> 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.</p> |
| spellingShingle | Duong, VT Tu, LTP Tuyen, HT Nhi, LTQ Campbell, JI Van Minh, P Le Phuc, H Chau, TTH Ngoc, NM Vi, LL Jenkins, C Okeke, I Higginson, E Baker, S 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 |
| title | 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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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