PCR methodology and applications for the detection of human fungal pathogens

ABSTRACT

Introduction: Polymerase chain reaction (PCR) has emerged as a promising technology for the rapid and reliable detection and identification of medical mycoses. Recent technological advancements – including microarray, multiplex PCR with magnetic resonance, and beacon probes – have mitigated the technical difficulties of performing nucleic amplification in fungi, thereby improving the sensitivity and specificity of PCR-based assays. In this paper, we examine current applications of PCR in the diagnosis of human fungal infections and look ahead to emerging techniques that may play a larger role in molecular diagnostics in the future.

Areas covered: This review includes a brief overview of the advantages and disadvantages of PCR using various clinical specimens, manual versus automated DNA extraction procedures, panfungal versus specific targets, and spectrum of pathogens detected. This is followed by a brief synopsis of species-specific PCR approaches and a more in-depth look at the obstacles to widespread implementation.

Expert commentary: The review concludes with a short perspective for the next five years, including the hurdles to standardization and validation, as well as the role of PCR coupled with electrospray-ionization mass spectrometry (PCR/ESI-MS) or nuclear magnetic resonance for the diagnosis of medical mycoses.

KEYWORDS:

• Polymerase chain reaction
• diagnostics
• amplicon
• dimorphic fungi
• mold
• yeast

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Declaration of interest

MW. McCarthy has received consulting from Allergan, Inc. TJ. Walsh receives research grants for experimental and clinical antimicrobial pharmacotherapeutics from Astellas, Cubist, Theravance, the Medicines Company, Allergan, Novartis, Merck and Pfizer. He has served as consultant to Astellas, Actavis, ContraFect, Drais, iCo, Novartis, Pfizer, Methylgene, SigmaTau and Trius. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

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Funding

This paper was not funded.