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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
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.
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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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