Saliva Diagnostics for Respiratory Diseases Using an Automated Integrated Platform

The ultimate goal of this study is to explore the possibility of utilizing saliva as a diagnostic fluid for respiratory diseases using an integrated automated platform based on an antibody-based multiplexed protein microarray.;Chapter 1 provides a summary of background knowledge related to this study, including discussions on inflammatory respiratory diseases---asthma and cystic fibrosis, saliva and saliva diagnostics, the six protein biomarkers selected for this study, protein immunoassays and protein microarrays, and the overall design for this study.;Chapter 2 describes the development of an antibody-based multiplexed protein microarray using fiber-optic bundles and polymer microspheres. The performance of the platform was characterized by recombinant human protein standards and blocking buffer. A total of 291 human saliva samples were also tested.;Chapters 3 and 4 analyze the results of saliva samples collected from patients with cystic fibrosis and asthma, respectively. A number of clinical features were collected from patients and healthy subjects for statistical analysis. Levels of proteins were assessed in different groups (patients with different respiratory diseases and healthy subjects) and subgroups (groups of patients with different clinical features) and were correlated with various clinical assessments.;Chapter 5 introduces the development of an automated integrated platform. For point-of-care diagnostics using human saliva for different respiratory diseases, an automated platform that can be used by personnel with minimal training was developed with our collaborators. For clinical applications, different parameters were optimized and the platform was characterized with recombinant human protein standards and blocking buffer.;Chapter 6 focuses on the clinical applications of the automated platform. To demonstrate its utility for point-of-care diagnostics, the platform was deployed at different clinics where hundreds of fresh saliva samples were collected and analyzed. The potential of utilizing these results to assist clinical diagnosis was also evaluated.;Chapter 7 discusses the future work for this study. The limitations of the current study, possible optimizations for this platform, and potential future applications are discussed.;As a whole, this thesis describes the development of an integrated automated platform that enables multiplexed salivary protein profiling for several clinical applications.