Core 2: Virology, Molecular, and Histology Core

摘要-病毒学、分子学和组织学核心（核心2） 众所周知，尽管严格遵守协议，但隐藏的或微妙的因素， 实验室可以使诸如关于病毒繁殖和qPCR的技术产生不同的结果， 成果。因此，为了确保跨项目的一致性，最大限度地减少混淆，并最大限度地提高结果的影响， 病毒学、分子学和组织学核心。这个核心将由彼得巴里博士领导，他带来了 在病毒原种繁殖和病毒参数分析方面， 感染（包括病毒载量、组织病理学和抗原表达的定量）。这些方法 是该项目的假设解决免疫（项目1）和病毒学的严格测试的核心 （项目2）先天性CMV感染（cCMV）的决定因素，并为NHP核心产生病毒储备， 纯化的DAN以评估病毒变体的出现/选择（核心3），以及病毒变体的可定量测量（核心3）。 复制和疾病进行严格的统计分析（核心4）。病毒学、分子学、 和组织学核心如下：RhCMV的嗜上皮性储液的良好表征的高滴度储液 (Aim 1）对接种动物的组织和体液进行灵敏度分析，以定量RhCMV基因组拷贝 通过PCR（qPCR）（目的2）和组织病理学/免疫组织化学（IHC）分析表征的数量 病毒病理学和抗原表达（目的3）。提供单一来源的病毒和敏感和 RhCMV基因组拷贝数和抗原表达的特异性测定将使起始材料一致 和分析技术，这些能力对评估本计划的总体目标至关重要： 定义影响cCMV传播的母体免疫应答和病毒特征， 胎儿感染的结果。

ABSTRACT – Virology, Molecular, and Histology Core (Core 2) It is well known that, despite strict adherence to protocols, hidden or subtle factors that vary among laboratories can cause techniques such as those concerning virus propagation and qPCR to produce differing outcomes. Thus, to ensure cross-project consistency, minimize confounding, and maximize impact of results, a Virology, Molecular, and Histology Core is proposed. This Core will be led by Dr. Peter Barry, who brings to the Program extant and well established techniques in virus stock propagation and analyses of viral parameters of infection (including quantification of viral loads, histopathology, and antigen expression). These approaches are central to rigorous testing of the project's hypotheses addressing the immunologic (Project 1) and virologic (Project 2) determinants of congenital CMV infection (cCMV), and generating virus stocks for the NHP Core, purified DAN to assess for emergence/selection of viral variants (Core 3), and quantifiable measures of viral replication and disease for rigorous statistical analyses (Core 4). Specific outputs of the Virology, Molecular, and Histology Core are as follows: well-characterized, high titer stocks of epithelial-tropic stocks of RhCMV (Aim 1), sensitive analyses of tissues and fluids from inoculated animals to quantify RhCMV genome copy numbers by PCR (qPCR) (Aim 2), and histopathologic/immunohistochemical (IHC) analyses to characterize viral pathology and antigen expression (Aim 3). Providing singly sourced stocks of virus and sensitive and specific assays for RhCMV genome copy numbers and antigen expression will enable uniform starting material and analytic techniques—capabilities that are critical for evaluating the overall objective of this Program: to define the maternal immune responses and viral characteristics that impact cCMV transmission and the outcome of fetal infection.