Plasma-based Detection of Valley Fever

Project: Research project

Project Details

Description

Plasma-based Detection of Valley Fever PLASMA-BASED DETECTION OF VALLEY FEVER Coccidioidomycosis or "Valley Fever" (VF) is caused by the fungus Coccidioides sp., which lives in soils of the southwestern US and San Juaquin Valley in California. When soil is disturbed, humans and animals can inhale fungal spores that germinate in the lung to produce a respiratory infection that ranges in severity from mild to life-threatening, and can easily be misdiagnosed as community acquired pneumonia. Because VF is caused by a fungus, it is important to differentiate VF patients from those who may have viral or bacterial pneumonia. Currently available tests to distinguish VF from other community acquired pneumonia, are based on the ability of patients to mount an antibody response to the fungus. Unfortunately, the antibody-based tests are inadequate for many patients because it may take weeks to months to develop such an antibody response (leading to delayed diagnosis), and many immunocompromised patients are unable to mount any antibody response at all. Therefore a blood test to detect coccidioidal proteins would allow a definitive diagnosis to be made even if serology is negative. Plasma is an ideal source to detect markers of infection because blood circulates through every organ, including lungs. The problem with plasma is that biologically informative molecules are obscured by large, highly abundant proteins in plasma (albumin, transferrin, etc.). To overcome this problem we subjected plasma to ultrafiltration followed by mass spectrometry (LC-MSIMS) to identify coccidioidal peptides in plasma from patients with VF. Preliminary data indicate that peptides from two coccidioidal proteins were identified and shared among at least two VF patients. The first goal of this pilot study is to determine the frequencies of coccidioidal peptides in patients with active Valley Fever. The second goal is to generate antibodies to each so that a rapid blood test (antigen capture ELISA) can be developed. Plasma-based Detection of Valley Fever One peptide was identified in plasma, common among 20 patients with active Coccidioidomycosis. A monoclonal antibody was generated against the peptide. The Mab detects a previously unknown parent protein from a Coccidioidal lysate in western blotting. We need to use the antibody to affinitypurify the parent antigen from fungal lysate followed by mass spec analysis of the parent protein. This will complete our studies and allow us to have a complete story to publish.
StatusFinished
Effective start/end date2/1/126/30/12

Funding

  • ASU: Mayo Seed Grant: $8,000.00

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