Date of Award

5-2016

Degree Type

Thesis

Degree Name

Master of Science

Department

Biology

Program

Biology

First Advisor/Chairperson

Josh Sharp

Second Advisor

Robert Belton

Third Advisor

Donna Becker

Abstract

Staphylococcus aureus is the most commonly isolated human associated bacterial pathogen. It plays an important role in skin and soft-tissue infections, pneumonia, endocarditis, osteomyelitis, foreign-body infections, and sepsis. S. aureus diagnosis and treatment requires a minimum of 24-48. With this in mind, previous studies suggest that faster pathogen identification has been linked to improved patient outcomes. Improved patient outcomes including a reduction in hospitalization time, decreased risk of nosocomial infections, and decreased in medical costs. The impact of faster identification on patient outcome has led us to develop an alternative method of S. aureus identification via ImmunoMagnetic Separation (IMS) and laser-light scattering identification technology. With this method, we hypothesized that anti-Protein A conjugated to magnetic DynaBeads could bind to surface Protein A on S. aureus from swab sample and facilitate their isolation upon exposure to a magnetic field within a 4-8 hour procedure. S. aureus cells isolated by IMS would then be accurately identified using laser-light scattering technology in less than 5 minutes. MIT identification accuracy analysis was conducted and displayed that both laboratory and clinical Staphylococcus species strains identified at a rate greater than 95% and negative control strains identified at a rate less than 1%. Our developed methods displayed statistically significant (P < 0.001) specificity for S. aureus and capture efficiency greater than 80%. The combination of IMS and laser-light identification gives a rapid and accurate identification in less than 8 hours, which is significantly less than traditional culture based identification methods.

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