Microbiology and Biospecimen Core

微生物学和生物学家核心(“MicroBio”核心)的建立是为了提供病毒和细菌 诊断、纯化的人鼻病毒(HRV)制剂以及培养的原代呼吸道和鼻腔上皮 三个拟议项目中的每一个都有单元格。核心将为本计划的项目II提供支持 更新应用程序，能够识别呼吸道样本中的所有常见呼吸道病毒 并确定HRV的个别种类/类型/毒株。所选细菌的数量 鼻腔连续感染致病菌(卡他莫拉氏菌、流感嗜血杆菌和肺炎链球菌) 灌洗液或痰标本将通过多重定量聚合酶链式反应进行检测，部分测序将用于 确定顺序检测到的细菌是代表单一菌株(定植)还是代表系列 不同菌株的感染。此外，该核心还将支持L三号工程的实验 提供(1)高滴度和均匀的HRV病毒库(2)针对特定HRV蛋白的抗体，(3) 原代呼吸道和鼻腔上皮细胞的培养，以及(4)执行标准化的HRV感染性 检测(空斑和TCID50)和定量RT-PCR；(5)感染鼻黏膜上皮细胞 心率变异性。除了实现这些目标外，微生物核心还将提供使用这些技术的专业知识 为海岸人员和其他合作人员提供试剂和病毒和细菌诊断分析 研究人员，并可为重新获得呼吸道病毒分子鉴定和 细菌、HRV的分子生物学和原代人类呼吸道上皮细胞的使用。

The Microbiology and Biospecimen Core ("MicroBio" Core) is established to provide viral and bacterial diagnostics, purified human rhinovirus (HRV) preparations, and cultured primary airway and nasal epithelial cells to each of the three proposed projects. The Core will provide support to Project II of this Program renewal application with the capability to identify all common respiratory viruses in samples of airway secretions and determine the individual species/type/strain of HRV. The quantity of selected bacterial pathogens {Moraxella catanrhalis, Hemophilus influenza and Streptococcus pneumoniae) in serial nasal lavage or sputum samples will be determined by multiplex qPCR, and partial sequencing will be used to determine whether bacteria that are sequentially detected represent a single strain (colonization) or serial infections with distinct strains. In addition, this Core will support the experiments of Projects l-lll by supplying (1) high-titer and uniform HRV virus stocks (2) antibodies targeting specific HRV proteins, (3) cultures of primary airway and nasal epithelial cells, and by (4) performing standardized HRV infectivity assays (plaque and TCID50) and quantitative RT-PCR and (5) infecting nasal epithelial cell cultures with HRV. In addition to fulfilling these aims, the MicroBio Core will also provide expertise for the use of these reagents and viral and bacterial diagnostic assays to COAST personnel and other collaborating investigators, and be available for consultation regaring molecular identification of respiratory viruses and bacteria, molecular biology of HRVs and the use of primary human airway epithelial cells.