PLoS ONE:抗药性HIV基因的成功鉴定

2010-12-12 00:00 · Duke

当前,全球范围内有3800万人感染了艾滋病毒,并且每年增加410万艾滋病患者。对于科学家而言,了解更多关于艾滋病病毒的变异和抗药性机制,这是长期有效治疗艾滋病所必不可少的。 来自维多利亚大学的生物医学工程师Stephanie Willerth发现了一项新的医学突破!它大大的加深了

当前,全球范围内有3800万人感染了艾滋病毒,并且每年增加410万艾滋病患者。对于科学家而言,了解更多关于艾滋病病毒的变异和抗药性机制,这是长期有效治疗艾滋病所必不可少的。

来自维多利亚大学的生物医学工程师Stephanie Willerth发现了一项新的医学突破!它大大的加深了人们对艾滋病的了解并且告诉我们如何去对待它!该文发表于PLoS ONE上。

“艾滋病病毒的变异频率非常高,这对于艾滋病患者而言糟糕透了,因为这些病毒最终对抗艾药产生了耐药性。”Willerth解释道。

Willerth及其搭档们对15000多种病毒展开了研究,有些病毒是以前从未研究过的。这为他们找到抗药性的特定基因提供了依据。显然,这也将最终可能帮助研究人员开发更有效的治疗艾滋病的药物。

研究者认为在研究过程中所使用的方法同样适用于一些难以治疗的疾病,比如猪流感、埃博拉病、流感甚至葡萄球菌感染。

“为了研究这些变异体,我们对这些病毒复制了无数次,特别是涉及到一些复杂的病毒,比如说HIV。”Willerth说,“绝大多数科学家都避免试图复制大量高危传染性病毒,但我们需要来复制大量的遗传物质。我们采用了一种独一无二的方法,我们分离到了HIV的遗传物质。当然在我们复制这些遗传物质时,病毒并不具备活性。”

Willerth及团队从那些长期患艾滋病感染者身上获得了病毒的小样本,当然,在这之前已经确认这些患者体内对艾滋病药物产生了抗药性的。研究者采用了新一代DNA测序技术来分析了这些遗传物质的组成。

生物谷推荐原文出处:

PLoS ONE 5(10): e13564. doi:10.1371/journal.pone.0013564

Development of a Low Bias Method for Characterizing Viral Populations Using Next Generation Sequencing Technology

Stephanie M. Willerth1#, Hélder A. M. Pedro2#, Lior Pachter3, Laurent M. Humeau4, Adam P. Arkin2*, David V. Schaffer1,2*

1 Department of Chemical Engineering and the Helen Wills Neuroscience Institute, University of California, Berkeley, California, United States of America, 2 Department of Bioengineering, University of California, Berkeley, California, United States of America, 3 Department of Mathematics and Molecular and Cell Biology, University of California, Berkeley, California, United States of America, 4 VIRxSYS Corporation, Gaithersburg, Maryland, United States of America

Background

With an estimated 38 million people worldwide currently infected with human immunodeficiency virus (HIV), and an additional 4.1 million people becoming infected each year, it is important to understand how this virus mutates and develops resistance in order to design successful therapies.

Methodology/Principal Findings

We report a novel experimental method for amplifying full-length HIV genomes without the use of sequence-specific primers for high throughput DNA sequencing, followed by assembly of full length viral genome sequences from the resulting large dataset. Illumina was chosen for sequencing due to its ability to provide greater coverage of the HIV genome compared to prior methods, allowing for more comprehensive characterization of the heterogeneity present in the HIV samples analyzed. Our novel amplification method in combination with Illumina sequencing was used to analyze two HIV populations: a homogenous HIV population based on the canonical NL4-3 strain and a heterogeneous viral population obtained from a HIV patient's infected T cells. In addition, the resulting sequence was analyzed using a new computational approach to obtain a consensus sequence and several metrics of diversity.

Significance

This study demonstrates how a lower bias amplification method in combination with next generation DNA sequencing provides in-depth, complete coverage of the HIV genome, enabling a stronger characterization of the quasispecies present in a clinically relevant HIV population as well as future study of how HIV mutates in response to a selective pressure.

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