کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6132977 | 1593451 | 2015 | 23 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Rapid next-generation sequencing of dengue, EV-A71 and RSV-A viruses
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ویروس شناسی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Rapid next-generation sequencing of dengue, EV-A71 and RSV-A viruses Rapid next-generation sequencing of dengue, EV-A71 and RSV-A viruses](/preview/png/6132977.png)
چکیده انگلیسی
Accurate characterisation of viral strains constitutes a crucial objective for the management of modern virus collections. Next-generation sequencing (NGS) provides technical solution for fast and cost-effective full genome sequencing. Here, we report protocols for rapid full-genome characterisation of RNA viruses of medical importance: dengue virus, enterovirus A71 and respiratory syncytial virus A, based on a specific amplification step followed by NGS-sequencing. A subset of full-length genome sequences representing the genetic diversity of each virus type was selected in GenBank and used to design primer sets allowing the amplification of the complete genome in 3-8 overlapping PCR fragments. The technique was used for characterising 53 strains (33 DENV, 8 EV-A71, 12 RSV-A) from various genotypes and origins. In a single assay, and in just 4 days, it provided for all strains an excellent genomic coverage (â¼99% including complete ORF for all strains) and accurate sequences with high number of reads per position (250-3500 on average). The elaboration of specific PCR-based full-genome sequencing protocols for diverse virus groups is likely to revolutionise the characterisation of viral isolates in modern collection, but also to contribute in the next future to the study of RNA viruses directly from biological samples.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Virological Methods - Volume 226, 15 December 2015, Pages 7-14
Journal: Journal of Virological Methods - Volume 226, 15 December 2015, Pages 7-14
نویسندگان
Cécile Baronti, Géraldine Piorkowski, Isabelle Leparc-Goffart, Xavier de Lamballerie, Audrey Dubot-Pérès,