Broad-Spectrum Molecular Detection of Fungal Nucleic Acids by PCR-Based Amplification Techniques
Over the past decade, the incidence of life-threatening invasive fungal infections has dramatically increased. Infections caused by hitherto rare and emerging fungal pathogens are associated with significant morbidity and mortality among immunocompromised patients. These observations render the coverage of a broad range of clinically relevant fungal pathogens highly important. The so-called panfungal or, perhaps more correctly, broad-range nucleic acid amplification techniques do not only facilitate sensitive detection of all clinically relevant fungal species but are also rapid and can be applied to analyses of any patient specimens. They have therefore become valuable diagnostic tools for sensitive screening of patients at risk of invasive fungal infections. This chapter summarizes the currently available molecular technologies employed in testing of a wide range of fungal pathogens, and provides a detailed workflow for patient screening by broad-spectrum nucleic acid amplification techniques.
Key wordsInvasive fungal infections Emerging fungal pathogens Panfungal PCR Molecular fungal diagnostics Broad-range fungal diagnostics
This work was performed within the FUNGITECT project supported by a grant from the European Commission (N° 602125) within the 7th Framework Programme (FP7).
- 13.El-Sayed ZA, Hasan ZE, Nasr RAR (2012) Real-time PCR in the early detection of invasive fungal infection in immunodeficient infants and children. Egypt J Pediatr Allergy Immunol 10:67–74Google Scholar
- 17.De Pauw B, Walsh TJ, Donnelly JP et al (2008) Revised definitions of invasive fungal disease from the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) Consensus Group. Clin Infect Dis 46:1813–1821CrossRefPubMedPubMedCentralGoogle Scholar
- 21.White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (ed) PCR protocols. A guide to methods and applications. pp 315–322Google Scholar
- 23.Babouee B, Goldenberger D, Elzi L et al (2013) Prospective study of a panfungal PCR assay followed by sequencing, for the detection of fungal DNA in normally sterile specimens in a clinical setting: a complementary tool in the diagnosis of invasive fungal disease? Clin Microbiol Infect 19:E354–E357CrossRefPubMedGoogle Scholar
- 24.Duval SM, Donnelly JP, Barnes R, Loeffler J (2008) PCR-based methods with aspergillosis as a model. J Invasive Fungal Infect 2:46–51Google Scholar