Core 2: Virology Core

摘要-病毒学核心（核心2，负责人：Ann Chahroudi博士，埃默里大学） 实现拟议计划的目标需要对SIV储层进行精确的定量和表征 和病毒动力学病毒学核心的目标是以严格的方式支持该计划的两个项目 通过提供最先进的病毒学工具，包括（i）定量PCR， 分析细胞相关的SHIV DNA和RNA以及血浆SHIV RNA，（ii）组合的细胞培养和PCR- 基于定量可复制病毒的比例的测定，和（iii）病毒序列分析， 深度测序和单基因组分析（SGA）。病毒学核心将由查赫鲁迪博士领导， 在埃默里大学的实验室里，她与杜克合作， 测序专业知识，为项目1和2提供标准化技术， 确定可解释和可推广的结果。核心小组提出以下具体目标：1） 血浆和细胞中病毒载量的定量PCR分析。2)水库规模估算， 复制能力/诱导型病毒的定量;和3）ART前和反弹的序列分析 与ART期间和反弹时的细胞相关SHIV变体相比，病毒血症。这些目标将利用 由Chahroudi博士的实验室开发的SIV/SHIV定量检测方法， 所用动物模型的灵敏度、特异性和重现性，以及SIV/HIV测序 杜克人类疫苗研究所的专家。病毒学核心将服务于这两个项目，并与 这两个行政和统计核心和非人类灵长类动物的核心，以实现我们的计划， 共同的目标。因此，病毒学核心在生成可靠和可量化的数据方面处于有利地位， 严格评估本计划中病毒反弹的起源、预测因素和病毒学相关因素。

ABSTRACT – Virology Core (Core 2, Leader: Dr. Ann Chahroudi, Emory University) Achieving the proposed Program's goal requires precise quantitation and characterization of SHIV reservoirs and viral dynamics. The Virology Core's objective is to support the Program's two Projects in the rigorous testing of their central hypotheses by providing state-of-the-art virological tools including (i) quantitative PCR analyses of cell-associated SHIV DNA and RNA and plasma SHIV RNA, (ii) combined cell culture and PCR- based assays quantifying the proportion of replication-competent viruses, and (iii) viral sequence analyses by deep sequencing and single genome analysis (SGA). The Virology Core will be led by Dr. Chahroudi, with both routine and novel assays performed in her laboratory at Emory University, and in collaboration with Duke sequencing expertise to provide standardized techniques across Projects 1 & 2 that will permit the ascertainment of interpretable and generalizable results. The Core proposes the following Specific Aims: 1) Quantitative PCR analysis of viral loads in plasma and cells. 2) Estimation of the reservoir size by quantification of replication-competent / inducible virus; and 3) Sequence analyses of pre-ART and rebound viremia in comparison to cell-associated SHIV variants during ART and at rebound. These Aims will make use SIV/SHIV quantification assays developed by Dr. Chahroudi's laboratory and rigorously evaluated for sensitivity, specificity and reproducibility in the animal model to be used, as well as SIV/HIV sequencing expertise of the Duke Human Vaccine Institute. The Virology Core will serve both Projects and interact with both the Administrative and Statistical Core and the Nonhuman Primate Core of this Program to achieve our shared goals. The Virology Core is thus strongly positioned to generate reliable and quantifiable data to rigorously assess the origin, predictors, and virologic correlates of viral rebound in this Program.