Clinical evaluation of a Mucorales-specific real-time PCR assay in tissue and serum samples

• Published in: Journal of medical microbiology Vol. 65; no. 12; pp. 1414 - 1421
• Main Authors: Springer, Jan, Lackner, Michaela, Ensinger, Christian, Risslegger, Brigitte, Morton, Charles Oliver, Nachbaur, David, Lass-Flörl, Cornelia, Einsele, Hermann, Heinz, Werner J, Loeffler, Juergen
• Format: Journal Article
• Language: English
• Published: England 01.12.2016
• Subjects: Adult; Aged; DNA Probes; DNA, Fungal - analysis; DNA, Fungal - biosynthesis; DNA, Fungal - genetics; Female; Fungemia - microbiology; Humans; Invasive Fungal Infections - diagnosis; Invasive Fungal Infections - microbiology; Male; Middle Aged; Molecular Diagnostic Techniques - methods; Mucorales - genetics; Mucorales - isolation & purification; Mucormycosis - diagnosis; Mucormycosis - microbiology; Paraffin Embedding; Real-Time Polymerase Chain Reaction - methods; Risk Factors; RNA, Ribosomal, 18S - genetics; Young Adult
• ISSN: 0022-2615; 1473-5644
• DOI: 10.1099/jmm.0.000375

Molecular diagnostic assays can accelerate the diagnosis of fungal infections and subsequently improve patient outcomes. In particular, the detection of infections due to Mucorales is still challenging for laboratories and physicians. The aim of this study was to evaluate a probe-based Mucorales-specific real-time PCR assay (Muc18S) using tissue and serum samples from patients suffering from invasive mucormycosis (IMM). This assay can detect a broad range of clinically relevant Mucorales species and can be used to complement existing diagnostic tests or to screen high-risk patients. An advantage of the Muc18S assay is that it exclusively detects Mucorales species allowing the diagnosis of Mucorales DNA without sequencing within a few hours. In paraffin-embedded tissue samples this PCR-based method allowed rapid identification of Mucorales in comparison with standard methods and showed 91 % sensitivity in the IMM tissue samples. We also evaluated serum samples, an easily accessible material, from patients at risk from IMM. Mucorales DNA was detected in all patients with probable/proven IMM (100 %) and in 29 % of the possible cases. Detection of IMM in serum could enable an earlier diagnosis (up to 21 days) than current methods including tissue samples, which were gained mainly post-mortem. A screening strategy for high-risk patients, which would enable targeted treatment to improve patient outcomes, is therefore possible.