-
Something wrong with this record ?
Determining the analytical specificity of PCR-based assays for the diagnosis of IA: What is Aspergillus
CO. Morton, PL. White, RA. Barnes, L. Klingspor, M. Cuenca-Estrella, K. Lagrou, S. Bretagne, W. Melchers, C. Mengoli, AM. Caliendo, M. Cogliati, Y. Debets-Ossenkopp, R. Gorton, F. Hagen, C. Halliday, P. Hamal, K. Harvey-Wood, K. Jaton, G....
Language English Country Great Britain
Document type Journal Article, Multicenter Study
PubMed
28339744
DOI
10.1093/mmy/myw093
Knihovny.cz E-resources
- MeSH
- Aspergillus classification genetics isolation & purification MeSH
- Molecular Diagnostic Techniques methods MeSH
- Invasive Pulmonary Aspergillosis diagnosis MeSH
- Humans MeSH
- Polymerase Chain Reaction methods MeSH
- Sensitivity and Specificity MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Multicenter Study MeSH
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
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
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 of Debrecen Medical and Health Science Center Debrecen Hungary
University of Padua Padua Italy
VU University Medical Centre Amsterdam The Netherlands
References provided by Crossref.org
- 000
- 00000naa a2200000 a 4500
- 001
- bmc18011314
- 003
- CZ-PrNML
- 005
- 20180417153219.0
- 007
- ta
- 008
- 180404s2017 xxk f 000 0|eng||
- 009
- AR
- 024 7_
- $a 10.1093/mmy/myw093 $2 doi
- 035 __
- $a (PubMed)28339744
- 040 __
- $a ABA008 $b cze $d ABA008 $e AACR2
- 041 0_
- $a eng
- 044 __
- $a xxk
- 100 1_
- $a Morton, C Oliver $u Western Sydney University, Sydney, Australia.
- 245 10
- $a Determining the analytical specificity of PCR-based assays for the diagnosis of IA: What is Aspergillus / $c CO. Morton, PL. White, RA. Barnes, L. Klingspor, M. Cuenca-Estrella, K. Lagrou, S. Bretagne, W. Melchers, C. Mengoli, AM. Caliendo, M. Cogliati, Y. Debets-Ossenkopp, R. Gorton, F. Hagen, C. Halliday, P. Hamal, K. Harvey-Wood, K. Jaton, G. Johnson, S. Kidd, M. Lengerova, C. Lass-Florl, C. Linton, L. Millon, CO. Morrissey, M. Paholcsek, AF. Talento, M. Ruhnke, B. Willinger, JP. Donnelly, J. Loeffler, . ,
- 520 9_
- $a 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.
- 650 _2
- $a Aspergillus $x klasifikace $x genetika $x izolace a purifikace $7 D001230
- 650 _2
- $a lidé $7 D006801
- 650 _2
- $a invazivní plicní aspergilóza $x diagnóza $7 D055744
- 650 _2
- $a diagnostické techniky molekulární $x metody $7 D025202
- 650 _2
- $a polymerázová řetězová reakce $x metody $7 D016133
- 650 _2
- $a senzitivita a specificita $7 D012680
- 655 _2
- $a časopisecké články $7 D016428
- 655 _2
- $a multicentrická studie $7 D016448
- 700 1_
- $a White, P Lewis $u Public Health Wales Microbiology Cardiff, UK.
- 700 1_
- $a Barnes, Rosemary A $u Cardiff University School of Medicine, Cardiff.
- 700 1_
- $a Klingspor, Lena $u Karolinska University Hospital, Stockholm, Sweden.
- 700 1_
- $a Cuenca-Estrella, Manuel $u Spanish National Centre for Microbiology, Instituto de Salud Carlos III, Madrid, Spain.
- 700 1_
- $a Lagrou, Katrien $u University Hospitals Leuven, Department of Laboratory Medicine and National Reference Center for Mycosis, Leuven, Belgium, Belgium.
- 700 1_
- $a Bretagne, Stéphane $u Paris Diderot, Sorbonne Paris Cité University, Faculty of Medicine, Paris, France.
- 700 1_
- $a Melchers, Willem $u Radboud University Medical Centre, Nijmegen, Netherlands.
- 700 1_
- $a Mengoli, Carlo $u University of Padua, Padua, Italy.
- 700 1_
- $a Caliendo, Angela M $u Department of Medicine, Alpert Medical School of Brown University, Providence, Rhode Island and Aspergillus Technology Consortium, USA.
- 700 1_
- $a Cogliati, Massimo $u Dip. Scienze Biomediche per la Salute, Università degli Studi di Milano, Milan, Italy.
- 700 1_
- $a Debets-Ossenkopp, Yvette $u VU University Medical Centre, Amsterdam. The Netherlands.
- 700 1_
- $a Gorton, Rebecca $u Royal Free Hospital, London, United Kingdom.
- 700 1_
- $a Hagen, Ferry $u Canisius-Wilhelmina Hospital, Nijmegen. The Netherlands.
- 700 1_
- $a Halliday, Catriona $u Clinical Mycology Reference Laboratory, Pathology West, Westmead, Australia.
- 700 1_
- $a Hamal, Petr $u Department of Microbiology, Faculty of Medicine and Dentistry, Palacky University Olomouc, Czech Republic.
- 700 1_
- $a Harvey-Wood, Kathleen $u Southern General Hospital, Glasgow, Scotland. UK.
- 700 1_
- $a Jaton, Katia $u Institute of Microbiology, University Hospital of Lausanne, Switzerland.
- 700 1_
- $a Johnson, Gemma $u Blizard Institute of Cell and Molecular Science, Queen Mary University of London, London, United Kingdom.
- 700 1_
- $a Kidd, Sarah $u National Mycology Reference Centre, SA Pathology, Adelaide. Australia.
- 700 1_
- $a Lengerova, Martina $u Department of Internal Medicine - Hematology and Oncology, University Hospital Brno, Brno, Czech Republic.
- 700 1_
- $a Lass-Florl, Cornelia $u Innsbruck Medical University, Innsbruck, Austria.
- 700 1_
- $a Linton, Chris $u UK Mycology Reference Lab, Public Health England, Bristol, United Kingdom.
- 700 1_
- $a Millon, Laurence $u Laboratoire de Parasitologie-Mycologie Centre Hospitalier Universitaire, Besançon, France.
- 700 1_
- $a Morrissey, C Orla $u Alfred Health and Monash University, Melbourne, Australia.
- 700 1_
- $a Paholcsek, Melinda $u University of Debrecen Medical and Health Science Center, Debrecen. Hungary.
- 700 1_
- $a Talento, Alida Fe $u Department of Clinical Microbiology, Trinity College, Dublin, Ireland.
- 700 1_
- $a Ruhnke, Markus $u Charité Medical School, University of Berlin, Berlin. Germany.
- 700 1_
- $a Willinger, Birgit $u Medical University of Vienna, Vienna, Austria.
- 700 1_
- $a Donnelly, J Peter $u Radboud University Medical Centre, Nijmegen, Netherlands.
- 700 1_
- $a Loeffler, Juergen $u Wuerzburg University, Wuerzburg, Germany.
- 700 1_
- $a ,
- 773 0_
- $w MED00005131 $t Medical mycology $x 1460-2709 $g Roč. 55, č. 4 (2017), s. 402-413
- 856 41
- $u https://pubmed.ncbi.nlm.nih.gov/28339744 $y Pubmed
- 910 __
- $a ABA008 $b sig $c sign $y a $z 0
- 990 __
- $a 20180404 $b ABA008
- 991 __
- $a 20180417153318 $b ABA008
- 999 __
- $a ok $b bmc $g 1288799 $s 1008126
- BAS __
- $a 3
- BAS __
- $a PreBMC
- BMC __
- $a 2017 $b 55 $c 4 $d 402-413 $i 1460-2709 $m Medical mycology $n Med Mycol $x MED00005131
- LZP __
- $a Pubmed-20180404
