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JMD 2006, Vol. 8, No. 3
Copyright © 2006 American Society for Investigative Pathology & Association for Molecular Pathology

A Consensus on Fungal Polymerase Chain Reaction Diagnosis?

A United Kingdom-Ireland Evaluation of Polymerase Chain Reaction Methods for Detection of Systemic Fungal Infections

P. Lewis White^*, Richard Barton, Malcolm Guiver, Christopher J. Linton, Steve Wilson^¶, Melvyn Smith^||, Beatriz L. Gomez^**, Michael J. Carr, Patrick T. Kimmitt, Shila Seaton^**, Kumar Rajakumar, Tessa Holyoake, Chris C. Kibbler^**, Elizabeth Johnson, Richard P. Hobson, Brian Jones^¶¶, Rosemary A. Barnes^* on behalf of the United Kingdom Fungal Polymerase Chain Reaction Consensus Group

From the Department of Medical Microbiology and National Public Health Service for Wales Cardiff, * University Hospital of Wales, Cardiff; the Mycology Reference Centre, Leeds General Infirmary, Leeds; Molecular Diagnostics, Northwest Health Protection Agency, Manchester Laboratory, Manchester Royal Infirmary, Manchester; the Mycology Reference Laboratory, Health Protection Agency Southwest, Bristol; Birmingham Health Protection Agency, ^¶ Birmingham Heartlands Hospital, Birmingham; Health Protection Agency London, ^|| London South Specialist Virology Centre, Kings College Hospital (Dulwich Site), London; the Department of Medical Microbiology, ** Royal Free Hospital, London; the Department of Clinical Microbiology, University of Dublin, Trinity College, Dublin; the Department of Clinical Microbiology, University Hospitals of Leicester, Leicester; and the Section of Experimental Haematology, Cancer Sciences, and Molecular Pathology and the Department of Medical Microbiology, ^¶¶ Glasgow Royal Infirmary, Glasgow, United Kingdom

The limitations of classical diagnostic methods for invasive fungal infections (IFIs) have led to the development of molecular techniques to aid in the detection of IFIs. Despite good published performance, interlaboratory reproduction of these assays is variable, and no consensus has been reached for an optimal method. This publication describes the first multicenter study of polymerase chain reaction methods, for the detection of Aspergillus and Candida species, currently used in the UK and Ireland by distribution and analysis of multiple specimen control panels. All three Candida methods were comparable, achieving a satisfactory level of detection (10 cfu), and the method of preference was dependent on the requirements of the particular laboratory. The results for the five Aspergillus assays were more variable, but two methods (2Asp and 4Asp) were superior (10¹ conidia). Formally, the overall performances of the two Aspergillus assays were comparable ( statistic = 0.77). However, on the Roche LightCycler, there was a clear sample-type effect that greatly reduced the detection limit of the 4Asp method when testing whole blood samples. Therefore, the preferred Aspergillus method relied on the amplification platform available to the user. This study represents the initial process to achieve a consensus method for the diagnosis of IFIs.

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