Microbiology and Biospecimen Core

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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.