Clinical evaluation of an in-house panfungal real-time PCR assay for the detection of fungal pathogens
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ORIGINAL PAPER
Clinical evaluation of an in‑house panfungal real‑time PCR assay for the detection of fungal pathogens Iris Camp1 · Gabriele Manhart1,2 · Claudia Schabereiter‑Gurtner1,3 · Kathrin Spettel1 · Brigitte Selitsch1 · Birgit Willinger1 Received: 31 October 2019 / Accepted: 24 January 2020 © The Author(s) 2020
Abstract Purpose Due to an increasing incidence of invasive fungal infections, the availability of reliable diagnostic tools for the fast detection of a wide spectrum of fungal pathogens is of vital importance. In this study, we aimed to conduct an extensive clinical evaluation of a recently published in-house panfungal PCR assay on samples from suspected invasive fungal infections. Methods Overall 265 clinical samples from 232 patients with suspected invasive fungal disease (96 deep airway samples, 60 sterile fluids, 50 tissue biopsies, and 59 blood samples) were included. All samples underwent standard culture-based diagnostics and were additionally analyzed with our panfungal PCR assay. Results Overall, 55.1% of agreement between culture and the panfungal PCR was observed; in 17% of all samples partial concordance was noted, while results between culture and our PCR assay were not in agreement in 27.9%. Our panfungal assay performed better in samples from normally sterile sites, while samples from the deep airways yielded the highest rate of discordant (39.6%) results. In two tissue and three blood samples an invasive pathogen was only detected by PCR while cultures remained negative. Conclusion In combination with routine methods, our panfungal PCR assay is a valuable diagnostic tool. Patients at risk for invasive fungal infections might profit from the reduced time to pathogen identification. Keywords Broadrange · Real-time PCR · Invasive fungal infections · ITS2 · Panfungal
Introduction Advances of modern medicine have led to the increased availability and use of immunosuppressant drugs, and an ever growing number of immunosuppressed patients. These patients are at high risk for invasive fungal infections, and infections with rare fungal pathogens are gaining relevance [1–11]. Thus, new diagnostic tools with the ability to detect common but also unusual and emerging fungal pathogens are urgently needed. Since disease progression—especially in immunocompromised patients—can be rapid, early * Birgit Willinger [email protected] 1
Division of Clinical Microbiology, Department of Laboratory Medicine, Medical University of Vienna, Vienna, Austria
2
Present Address: Institute for Medical Biochemistry, University of Veterinary Medicine Vienna, Vienna, Austria
3
Present Address: Ingenetix GmbH, Vienna, Austria
detection is important for a favourable clinical outcome [12, 13]. Currently, culture methods and histology still are considered to be the gold standard for the diagnosis of fungal infections. However, histology often does not allow the discrimination between distinct fungal species, and—due to the slow growth of most fungal pathogens—culture methods ca
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