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A universal genome sequencing method for rotavirus A from human fecal samples which identifies segment reassortment and multi-genotype mixed infection
BACKGROUND: Genomic characterization of rotavirus (RoV) has not been adopted at large-scale due to the complexity of obtaining sequences for all 11 segments, particularly when feces are used as starting material. METHODS: To overcome these limitations, we developed a novel RoV capture and genome seq...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5404283/ https://www.ncbi.nlm.nih.gov/pubmed/28438140 http://dx.doi.org/10.1186/s12864-017-3714-6 |
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author | Dung, Tran Thi Ngoc Duy, Pham Thanh Sessions, October M. Sangumathi, Uma K. Phat, Voong Vinh Tam, Pham Thi Thanh To, Nguyen Thi Nguyen Phuc, Tran My Hong Chau, Tran Thi Chau, Nguyen Ngoc Minh Minh, Ngoc Nguyen Thwaites, Guy E. Rabaa, Maia A. Baker, Stephen |
author_facet | Dung, Tran Thi Ngoc Duy, Pham Thanh Sessions, October M. Sangumathi, Uma K. Phat, Voong Vinh Tam, Pham Thi Thanh To, Nguyen Thi Nguyen Phuc, Tran My Hong Chau, Tran Thi Chau, Nguyen Ngoc Minh Minh, Ngoc Nguyen Thwaites, Guy E. Rabaa, Maia A. Baker, Stephen |
author_sort | Dung, Tran Thi Ngoc |
collection | PubMed |
description | BACKGROUND: Genomic characterization of rotavirus (RoV) has not been adopted at large-scale due to the complexity of obtaining sequences for all 11 segments, particularly when feces are used as starting material. METHODS: To overcome these limitations, we developed a novel RoV capture and genome sequencing method combining commercial enzyme immunoassay plates and a set of routinely used reagents. RESULTS: Our approach had a 100% success rate, producing >90% genome coverage for diverse RoV present in fecal samples (Ct < 30). CONCLUSIONS: This method provides a novel, reproducible and comparatively simple approach for genomic RoV characterization and could be scaled-up for use in global RoV surveillance systems. TRIAL REGISTRATION (PROSPECTIVELY REGISTERED): Current Controlled Trials ISRCTN88101063. Date of registration: 14/06/2012 ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-017-3714-6) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5404283 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-54042832017-04-27 A universal genome sequencing method for rotavirus A from human fecal samples which identifies segment reassortment and multi-genotype mixed infection Dung, Tran Thi Ngoc Duy, Pham Thanh Sessions, October M. Sangumathi, Uma K. Phat, Voong Vinh Tam, Pham Thi Thanh To, Nguyen Thi Nguyen Phuc, Tran My Hong Chau, Tran Thi Chau, Nguyen Ngoc Minh Minh, Ngoc Nguyen Thwaites, Guy E. Rabaa, Maia A. Baker, Stephen BMC Genomics Methodology Article BACKGROUND: Genomic characterization of rotavirus (RoV) has not been adopted at large-scale due to the complexity of obtaining sequences for all 11 segments, particularly when feces are used as starting material. METHODS: To overcome these limitations, we developed a novel RoV capture and genome sequencing method combining commercial enzyme immunoassay plates and a set of routinely used reagents. RESULTS: Our approach had a 100% success rate, producing >90% genome coverage for diverse RoV present in fecal samples (Ct < 30). CONCLUSIONS: This method provides a novel, reproducible and comparatively simple approach for genomic RoV characterization and could be scaled-up for use in global RoV surveillance systems. TRIAL REGISTRATION (PROSPECTIVELY REGISTERED): Current Controlled Trials ISRCTN88101063. Date of registration: 14/06/2012 ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-017-3714-6) contains supplementary material, which is available to authorized users. BioMed Central 2017-04-24 /pmc/articles/PMC5404283/ /pubmed/28438140 http://dx.doi.org/10.1186/s12864-017-3714-6 Text en © The Author(s). 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Methodology Article Dung, Tran Thi Ngoc Duy, Pham Thanh Sessions, October M. Sangumathi, Uma K. Phat, Voong Vinh Tam, Pham Thi Thanh To, Nguyen Thi Nguyen Phuc, Tran My Hong Chau, Tran Thi Chau, Nguyen Ngoc Minh Minh, Ngoc Nguyen Thwaites, Guy E. Rabaa, Maia A. Baker, Stephen A universal genome sequencing method for rotavirus A from human fecal samples which identifies segment reassortment and multi-genotype mixed infection |
title | A universal genome sequencing method for rotavirus A from human fecal samples which identifies segment reassortment and multi-genotype mixed infection |
title_full | A universal genome sequencing method for rotavirus A from human fecal samples which identifies segment reassortment and multi-genotype mixed infection |
title_fullStr | A universal genome sequencing method for rotavirus A from human fecal samples which identifies segment reassortment and multi-genotype mixed infection |
title_full_unstemmed | A universal genome sequencing method for rotavirus A from human fecal samples which identifies segment reassortment and multi-genotype mixed infection |
title_short | A universal genome sequencing method for rotavirus A from human fecal samples which identifies segment reassortment and multi-genotype mixed infection |
title_sort | universal genome sequencing method for rotavirus a from human fecal samples which identifies segment reassortment and multi-genotype mixed infection |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5404283/ https://www.ncbi.nlm.nih.gov/pubmed/28438140 http://dx.doi.org/10.1186/s12864-017-3714-6 |
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