Coccidioidomycosis, more commonly known as Valley Fever, is a disease caused by pulmonary exposure to Coccidioides spp. fungi. This dangerous pathogen is rampant in the southwestern United States, and its complex nature has made it difficult to thoroughly understand the effects of Coccidioides on the human body, particularly diagnosis of the disease has proven challenging and time-consuming. Currently, it is not known what components of fungal endospores and spherules growing in the lung circulate in blood. In an attempt to identify protein components of Coccidioides in the blood of those infected, this ongoing study aims to extract and isolate the Coccidioides proteins from the plasma of infected human subjects, identify them via mass spectrometry, and validate them as biomarkers of exposure for more rapid Valley Fever diagnosis. Plasma samples of infected and healthy patients were purified by affinity chromatography using lectin that has been shown to bind to spherules and endospores in infected lung tissues. Fungal proteins are eluted from the lectin, separated, and trypsin-digested for qualitative analysis by matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF MS). Results obtained thus far provide mass spectrometric evidence for the ability to extract and identify Coccidioides proteins in blood plasma using this method. This study is one of multiple steps required to bring us closer to a rapid, accurate method for diagnosing coccidioidomycosis enabled here by mass spectrometry-based proteomics.

Development of a proteomic-based analytical method for the isolation and identification of <i>Coccidioides</i> spp. proteins detectable in human blood plasma

248th National Meeting (2014)

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Development of a proteomic-based analytical method for the isolation and identification of Coccidioides spp. proteins detectable in human blood plasma