Name: amplifying and quantifying hiv-1 rna in hiv infected individuals with viral loads below the limit of detection by standard clinical assays
Uploaded: 2011-09-26T12:00:00
Description: Watch this Scientific Journal Video about Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays at JoVE.com

作者要感谢的患者参加的HIV - 1的许多研究。 江铃汽车的FM柯比基金会的支持下与美国癌症协会的研究教授。

1。从大量的血浆提取的RNA 在图1和2提供了一个单拷贝检测（SCA）的概述和单基因组测序（SGS）的协议。 <ol><li>要获得7毫升血浆，旋转约14毫升的血液收集在15毫升EDTA，15分钟，在2600 XG（肝素）无制动收集管。移液器（一定要避免巴菲涂层）为15毫升管的血浆。 </li><li>如果RNA是单拷贝检测（SCA），秒杀Rous肉瘤病毒内部的病毒粒子控制（RCAS）30000份血浆中提取。 RCAS病毒前准?...

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艾滋病毒1感染者抗逆转录病毒药物联合治疗（车）或自然控制感染病毒载量非常低，通常约1副本每毫升血浆4，11，12，17，18。自然控制患者的病毒载量，往往个人设置1，2，17点左右波动。检测的灵敏度，此处所描述的高度依赖的血浆输入量。一般来说，我们已与7毫升血浆，但取得了积极的成果，可以从少量的血浆，以及根据实际血浆病毒载量。此处所描述的RNA提取方法“，这?...

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amplifying and quantifying hiv-1 rna in hiv infected individuals with viral loads below the limit of detection by standard clinical assays

，获得患者的洞察力病毒动态和病毒宿主相互作用，病毒基因放大和量化，在HIV - 1病毒载量低于检测标准检测的限制（低于50-75拷贝/ ml）感染者HIV - 1 RNA是必要的日志自然控制的感染和抗逆转录病毒联合疗法（CART）的那些。 在这里，我们介绍了如何由单一的基因组测序（SGS协议）13，19和如何准确地量化在与病毒载量低（单拷贝检测（SCA）协议）12，20的患者的HIV - 1 RNA病毒的基因组扩增。 单拷贝检测的实时PCR检测的灵敏度取决于被检测血浆量。如果一个单一的病毒基因组是7毫升血浆中检测到，然后报道RNA拷贝数为0.3拷贝/ ml。该检测RNA提取效率的内部控制测试，并可能扩增DNA或污染控制。测量患者样本一式三份。 ，现在已经广泛使用，被视为非劳动密集的3，7，12，14，15的单基因组测序法（SGS）的，是一个限制的稀释检测，其中端点cDNA产物稀释96孔蔓延板。根据泊松分布，不到1 / 3的水井时给予的产品，有一个80％的机会的PCR产物被放大的结果，从一个单一的基因分子。 SGS已超过克隆没有受到重采样和PCR引入重组19偏颇的优势。然而，SCA和SGS的扩增成功取决于引物的设计。两种检测为HIV - 1 B亚型，但可以适应不断变化的引物，探针和标准的其他亚型和基因组的其他地区。

Watch this Scientific Journal Video about Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays at JoVE.com

Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays

量化血浆中HIV - 1 RNA水平和测序单一的HIV - 1病毒载量低于检测限（50-75拷贝/ ml）与个人的基因组是困难的。在这里，我们描述了如何提取和量化使用实时PCR检测方法能够可靠措施，HIV - 1 RNA下降到0.3拷贝/ ml，以及如何放大由单一的基因组测序的病毒基因组病毒载量非常低的样品，血浆病毒RNA。

在艾滋病毒感染者病毒载量低于检测限制标准的临床实验放大和量化HIV - 1 RNA

Amplifying viral genes and quantifying HIV-1 RNA in HIV-1 infected individuals with viral loads below the limit of detection by standard assays (below ...

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Prediction of HIV-1 Coreceptor Usage Tropism by Sequence Analysis using a Genotypic Approach

使用基因型的方法预测HIV-1辅助用途（归经）的序列分析

Maraviroc (MVC) is the first licensed antiretroviral drug from the class of coreceptor antagonists. It binds to the host coreceptor CCR5, which is used by ...

In vitro Uncoating of HIV-1 Cores

In vitro Uncoating of HIV-1 Cores

在体外对HIV - 1核心脱壳

The genome of the retroviruses is encased in a capsid surrounded by a lipid envelope. For lentiviruses, such as HIV-1, the conical capsid shell is ...

Imaging of HIV-1 Envelope-induced Virological Synapse and Signaling on Synthetic Lipid Bilayers

艾滋病毒的成像1信封引起的病毒学突触信号合成脂双层

Human immunodeficiency virus type 1 (HIV-1) infection occurs most efficiently via cell to cell transmission2,10,11. This cell to cell transfer between ...

New Tools to Expand Regulatory T Cells from HIV-1-infected Individuals

新工具展开调节性T细胞的HIV-1感染的个体

CD4+ Regulatory T cells (Tregs) are potent immune modulators and serve an important function in human immune homeostasis. Depletion of Tregs has led to ...

Assessing the Innate Sensing of HIV-1 Infected CD4+ T Cells by Plasmacytoid Dendritic Cells Using an Ex vivo Co-culture System.

Assessing the Innate Sensing of HIV-1 Infected CD4+ T Cells by Plasmacytoid Dendritic Cells Using an Ex vivo Co-culture System.

评估的HIV-1感染的CD4的先天传感<sup&gt; +</sup&gt;性T细胞浆细胞样树突细胞使用<em&gt;体外</em&gt;共培养体系。

HIV-1 innate sensing requires direct contact of infected CD4+ T cells with plasmacytoid dendritic cells (pDCs). In order to study this process, the ...

Nucleocapsid Annealing-Mediated Electrophoresis (NAME) Assay Allows the Rapid Identification of HIV-1 Nucleocapsid Inhibitors

Nucleocapsid Annealing-Mediated Electrophoresis NAME Assay Allows the Rapid Identification of HIV-1 Nucleocapsid Inhibitors

核衣壳退火介导电泳（NAME）分析允许HIV-1核衣壳抑制剂的快速鉴定

RNA or DNA folded in stable tridimensional folding are interesting targets in the development of antitumor or antiviral drugs. In the case of HIV-1, viral ...

Isolation of Exosomes from the Plasma of HIV-1 Positive Individuals

从HIV-1阳性者的血浆分离外来体的

Exosomes are small vesicles ranging in size from 30 nm to 100 nm that are released both constitutively and upon stimulation from a variety of cell types. ...

Phagosome Migration and Velocity Measured in Live Primary Human Macrophages Infected with HIV-1

吞噬体偏移和速度与HIV-1感染的活小学的人巨噬细胞测量

Macrophages are phagocytic cells that play a major role at the crossroads between innate and specific immunity. They can be infected by the human ...