Determining the analytical specificity of PCR-based assays for the diagnosis of IA: What is Aspergillus?
| Autor: | Massimo Cogliati, Yvette J. Debets-Ossenkopp, Juergen Loeffler, Birgit Willinger, Petr Hamal, Catriona Halliday, Angela M. Caliendo, Rosemary Ann Barnes, Martina Lengerová, C. Orla Morrissey, Melinda Paholcsek, Katia Jaton, Katrien Lagrou, Willem J. G. Melchers, Lena Klingspor, Kathleen Harvey-Wood, Laurence Millon, Gemma Johnson, Cornelia Lass-Flörl, Carlo Mengoli, Stéphane Bretagne, Sarah E. Kidd, J. Peter Donnelly, Markus Ruhnke, Alida Fe Talento, Rebecca Gorton, Christopher J. Linton, P. Lewis White, Ferry Hagen, Manuel Cuenca-Estrella, C. Oliver Morton |
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| Přispěvatelé: | Western Sydney University, Public Health Wales Microbiology Cardiff, School of Medicine [Cardiff], Institute of Medical Genetics [Cardiff]-Cardiff University, Karolinska University Hospital [Stockholm], Instituto de Salud Carlos III [Madrid] (ISC), Université Catholique de Louvain = Catholic University of Louvain (UCL), Université Paris Diderot, Sorbonne Paris Cité, Paris, France, Université Paris Diderot - Paris 7 (UPD7), Radboud University Medical Center [Nijmegen], Regional Reference Centre of Infectious Diseases,Microbiology and Virology Unit,University of Padova, Warren Alpert Medical School of Brown University, Università degli Studi di Milano [Milano] (UNIMI), Neuroscience Amsterdam, VU University Medical Centre, 1081HV 1117, Amsterdam, Royal Free Hospital, London, Canisius-Wilhelmina Hospital [Nijmegen, The Netherlands], Clinical Mycology Reference Laboratory, Pathology West, Westmead, Australia, Palacky University Olomouc, Southern General Hospital, Glasgow, University of Lausanne and University Hospital Centre, School of Biological and Chemical Sciences, Queen Mary University of London, National Mycology Reference Centre, SA Pathology, Adelaide, University Hospital Brno, Innsbruck Medical University [Austria] (IMU), Public Health England [London], Laboratoire Chrono-environnement - CNRS - UBFC (UMR 6249) (LCE), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Alfred Health, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary, Trinity College Dublin, Abt. Onkologie und Hämatologie, Med. Klinik u. Poliklinik II, Charité - UniversitätsMedizin = Charité - University Hospital [Berlin], Medizinische Universität Wien = Medical University of Vienna, University of Würzburg = Universität Würzburg |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
030106 microbiology Klinikai orvostudományok Aspergillosis medicine.disease_cause Polymerase Chain Reaction Sensitivity and Specificity Cross-reactivity Microbiology law.invention Aspergillus PCR 03 medical and health sciences law Biopsy medicine Humans Gene Polymerase chain reaction [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology Invasive Pulmonary Aspergillosis Aspergillus biology medicine.diagnostic_test Orvostudományok General Medicine Assay sensitivity Ribosomal RNA biology.organism_classification medicine.disease analytical specificity Women's cancers Radboud Institute for Health Sciences [Radboudumc 17] Infectious Diseases Molecular Diagnostic Techniques cross reactivity detection range Inflammatory diseases Radboud Institute for Molecular Life Sciences [Radboudumc 5] |
| Zdroj: | Medical Mycology, 55, 4, pp. 402-413 Medical Mycology Medical Mycology, Oxford University Press, 2016, 55 (4), pp.402-413. ⟨10.1093/mmy/myw093⟩ Medical Mycology, 55, 402-413 |
| ISSN: | 1369-3786 1365-280X |
| Popis: | Contains fulltext : 175113.pdf (Publisher’s version ) (Closed access) 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. |
| Databáze: | OpenAIRE |
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