A universal genome sequencing method for rotavirus A from human fecal samples which identifies segment reassortment and multi-genotype mixed infection.
A universal genome sequencing method for rotavirus A from human fecal samples which identifies segment reassortment and multi-genotype mixed infection.
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DOI：
10.1186/s12864-017-3714-6
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发表时间：
2017-04-24
期刊：
影响因子：
4.4
通讯作者：
Baker S
中科院分区：
文献类型：
--
作者：
Dung TTN;Duy PT;Sessions OM;Sangumathi UK;Phat VV;Tam PTT;To NTN;Phuc TM;Hong Chau TT;Chau NNM;Minh NN;Thwaites GE;Rabaa MA;Baker S
轮状病毒(ROV)的基因组特征尚未大规模采用，因为获得所有11个片段的序列很复杂，特别是当以粪便为起始材料时。为了克服这些局限性，我们开发了一种新的ROV捕获和基因组测序方法，该方法结合了商用酶免疫分析平板和一系列常规使用的试剂。我们的方法有100%的成功率，对粪便样本中存在的各种ROV产生了90%的基因组覆盖率(CT <； 30)。这种方法提供了一种新颖、可重复性和相对简单的基因组ROV表征方法，并可扩大用于全球ROV监视系统。当前对照试验ISRCTN88101063。注册日期：2012年6月14日本文的在线版本(doi：10.1186/s12864-017-3714-6)载有补充材料，可供授权用户使用。
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. 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. Our approach had a 100% success rate, producing >90% genome coverage for diverse RoV present in fecal samples (Ct < 30). 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. Current Controlled Trials ISRCTN88101063. Date of registration: 14/06/2012 The online version of this article (doi:10.1186/s12864-017-3714-6) contains supplementary material, which is available to authorized users.