Microbiology and Biospecimen Core

建立微生物学和生物标本核心（“微生物”核心），以提供病毒和细菌 诊断、纯化的人鼻病毒（HRV）制剂和培养的原代气道和鼻上皮 三个项目中的每一个项目。核心将为本计划的项目II提供支持 更新应用程序，能够识别气道样本中的所有常见呼吸道病毒 分泌物，并确定个别物种/类型/应变的HRV。选择的细菌数量 病原体（卡他莫拉菌、流感嗜血杆菌和肺炎链球菌） 灌洗液或痰液样本将通过多重qPCR测定，部分测序将用于 确定连续检测到的细菌是否代表单一菌株（定殖）或系列菌株 不同菌株的感染。此外，该核心将通过以下方式支持项目I-III的实验： 提供（1）高滴度和均匀的HRV病毒原液（2）靶向特异性HRV蛋白的抗体，（3） 原代气道和鼻上皮细胞的培养，以及通过（4）进行标准化的HRV感染性 测定（噬斑和TCID 50）和定量RT-PCR，和（5）用 心率变异性除了实现这些目标，微生物核心还将提供使用这些技术的专业知识。 试剂和病毒和细菌诊断分析，以海岸人员和其他合作 研究人员，并可就呼吸道病毒的分子鉴定进行咨询， 细菌、HRV的分子生物学和原代人气道上皮细胞的使用。

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.