A wide array of PCR tests has been developed to aid the diagnosis of invasive aspergillosis (IA), providing technical diversity but limiting standardisation and acceptance. Methodological recommendations for testing blood samples using PCR exist, based on achieving optimal assay sensitivity to help exclude IA. Conversely, when testing more invasive samples (BAL, biopsy, CSF) emphasis is placed on confirming disease, so analytical specificity is paramount. This multicenter study examined the analytical specificity of PCR methods for detecting IA by blind testing a panel of DNA extracted from a various fungal species to explore the range of Aspergillus species that could be detected, but also potential cross reactivity with other fungal species. Positivity rates were calculated and regression analysis was performed to determine any associations between technical specifications and performance. The accuracy of Aspergillus genus specific assays was 71.8%, significantly greater (P < .0001) than assays specific for individual Aspergillus species (47.2%). For genus specific assays the most often missed species were A. lentulus (25.0%), A. versicolor (24.1%), A. terreus (16.1%), A. flavus (15.2%), A. niger (13.4%), and A. fumigatus (6.2%). There was a significant positive association between accuracy and using an Aspergillus genus PCR assay targeting the rRNA genes (P = .0011). Conversely, there was a significant association between rRNA PCR targets and false positivity (P = .0032). To conclude current Aspergillus PCR assays are better suited for detecting A. fumigatus, with inferior detection of most other Aspergillus species. The use of an Aspergillus genus specific PCR assay targeting the rRNA genes is preferential.

Alfred Health and Monash University Melbourne Australia

Blizard Institute of Cell and Molecular Science Queen Mary University of London London United Kingdom

Canisius Wilhelmina Hospital Nijmegen The Netherlands

Cardiff University School of Medicine Cardiff

Charité Medical School University of Berlin Berlin Germany

Clinical Mycology Reference Laboratory Pathology West Westmead Australia

Department of Clinical Microbiology Trinity College Dublin Ireland

Department of Internal Medicine Hematology and Oncology University Hospital Brno Brno Czech Republic

Department of Medicine Alpert Medical School of Brown University Providence Rhode Island and Aspergillus Technology Consortium USA

Department of Microbiology Faculty of Medicine and Dentistry Palacky University Olomouc Czech Republic

Dip Scienze Biomediche per la Salute Università degli Studi di Milano Milan Italy

Innsbruck Medical University Innsbruck Austria

Institute of Microbiology University Hospital of Lausanne Switzerland

Karolinska University Hospital Stockholm Sweden

Laboratoire de Parasitologie Mycologie Centre Hospitalier Universitaire Besançon France

Medical University of Vienna Vienna Austria

National Mycology Reference Centre SA Pathology Adelaide Australia

Paris Diderot Sorbonne Paris Cité University Faculty of Medicine Paris France

Public Health Wales Microbiology Cardiff UK

Radboud University Medical Centre Nijmegen Netherlands

Royal Free Hospital London United Kingdom

Southern General Hospital Glasgow Scotland UK

Spanish National Centre for Microbiology Instituto de Salud Carlos 3 Madrid Spain

UK Mycology Reference Lab Public Health England Bristol United Kingdom

University Hospitals Leuven Department of Laboratory Medicine and National Reference Center for Mycosis Leuven Belgium Belgium

University of Debrecen Medical and Health Science Center Debrecen Hungary

University of Padua Padua Italy

VU University Medical Centre Amsterdam The Netherlands

Western Sydney University Sydney Australia

Wuerzburg University Wuerzburg Germany

Citace poskytuje Crossref.org

An Overview of Systematic Reviews of Polymerase Chain Reaction (PCR) for the Diagnosis of Invasive Aspergillosis in Immunocompromised People: A Report of the Fungal PCR Initiative (FPCRI)-An ISHAM Working Group