Fungal Molecular Diagnostics

Utility of Coccidioides PCR Testing in the Clinical Setting⁴

February 2011

Since we now have several years of experience utilizing the Coccidioides real-time PCR assay, we have been able to do several studies evaluating the clinical utility of this approach. Two manuscripts have been recently accepted and are currently in press in the journal Mycopathologia. I have provided the reference information if you would like to learn more. In the first study, we reviewed the medical charts of 145 patients with suspected coccidiomycosis and found that the real-time PCR assay was superior to fungal culture for the diagnosis of pulmonary disease, but I should caution that the "n" was small for this study. The PCR assay was also found to be highly useful in rapidly ruling out coccidiomycosis with a greater than 99% negative predictive value.

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PCR Testing in the Clinical Setting4

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Jump to section:

• Introduction
• Objectives
• Traditional Methods of Fungal Diagnostics
• Traditional Methods
• Potential Advantages of Fungal Molecular Diagnostics
• Currently Available Molecular Diagnostics for Fungi
• Nucleic Acid Hybridization Probes³
• Nucleic Acid Hybridization Probes
• Nucleic Acid Hybridization Probes for Fungi
• Nucleic Acid Hybridization Probes
• DNA Sequencing for Fungi
• Ribosomal Structure: Fungi
• Traditional Sequencing Workflow
• Sequence Analysis
• Current Applications of D2 Fungal Sequencing at the Mayo Clinic
• Rapid, Direct Identification of Yeast from Blood Cultures
• PNA-FISH Basics
• PNA FISH for Candida spp—Direct Detection from Blood
• Yeast Identification Timeline from Positive Blood Bottle
• Potential Advantages of Direct Detection
• Fungal PCR Assays at Mayo Clinic
• Real-time PCR Workflow
• Pneumocystis PCR
• Pneumocystis LC-PCR Melt Curves
• Concordance of the Pneumocystis LightCycler PCR Assay and Calcofluor White Stain for BAL Fluid
• Coccidioides species Real-time PCR
• Melt-curve Analysis of Coccidioides spp Grown in Culture
• Utility of Coccidioides PCR Testing in the Clinical Setting⁴
• Meningeal Coccidioidomycosis Diagnosed by Real-Time PCR Analysis of Cerebrospinal Fluid⁵
• Histoplasma capsulatum/Blastomyces dermatitidis Real-Time PCR
• What lies in the future for fungal molecular diagnostics?
• Mass Spectrometric Identification Of Fungi
• Summary
• References
• Questions?
• Disclosure