Article (Scientific journals)
Direct detection of Aspergillus and azole resistance of Aspergillus fumigatus on broncho-alveolar lavage fluid. Validation of a new Aspergillus real-time PCR
Chong, Ga-Lai; Van de Sande, Wendy; Dingemans, Gijs et al.
2015In Journal of Clinical Microbiology
Peer Reviewed verified by ORBi
 

Files


Full Text
JCM.03216-14.full.pdf
Author preprint (1 MB)
Download

All documents in ORBi are protected by a user license.

Send to



Details



Keywords :
Aspergillus species; Aspergillus fumigatus; invasive aspergillosis; haematology; intensive care unit; polymerase chain reaction
Abstract :
[en] Introduction Azole resistance in Aspergillus fumigatus is increasingly reported. We describe the validation of AsperGenius® , a new multiplex real-time polymerase chain reaction (PCR) assay consisting of two multiplex real-time PCRs: one which identifies the clinically relevant Aspergillus species, and one which detects the TR34, L98H, T289A, Y121F mutations in CYP51A and differentiates susceptible from resistant A. fumigatus strains. Methods The diagnostic performance was tested on 37 bronchoalveolar lavage (BAL) samples from haematology patients and on 40 BAL samples from intensive care unit (ICU) patients using BAL galactomannan ≥1.0 or positive culture as the gold standard for the presence of Aspergillus. Results In the haematology and ICU groups combined, there were 22 BAL samples with IA (2 proven, 9 probable and 11 non-classifiable). Nineteen of the 22 BAL samples were positive according to the gold standard. The optimal cycle threshold value for the presence of Aspergillus was <36. Sixteen of the 19 BAL samples had a positive PCR (2 Aspergillus species and 14 A. fumigatus). This resulted in a sensitivity, specificity, positive and negative predictive value of 88.9%, 89.3%, 72.7% and 96.2% for the haematology group and 80.0%, 93.3%, 80.0% and 93.3% in the ICU group, respectively. The CYP51A real-time PCR confirmed 12 wildtype and 2 resistant strains (1 TR34/L98H and 1 TR46/Y121F/T289A mutant). Conclusion The AsperGenius® multiplex real-time PCR allows for a sensitive and fast detection of Aspergillus species directly in BAL samples. More importantly, this assay detects and differentiates wildtype from resistant strains even if BAL cultures remained negative.
Disciplines :
Microbiology
Author, co-author :
Chong, Ga-Lai;  Erasmus Medical Centre, Rotterdam > Department of internal medecine > Infectious diseases
Van de Sande, Wendy;  Erasmus Medical Centre > Medical Microbiology and infectious diseases
Dingemans, Gijs;  PathoNostics B.V. Maastricht, The Nederlands
Gaajetaan, Giel;  PathoNostics B.V. Maastricht, The Nederlands
Vonk, Alieke;  Erasmus Medical Centre > Medical Microbiology and infectious diseases
Hayette, Marie-Pierre ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Bactériologie, mycologie, parasitologie, virologie
Van Tegelen, Dennis;  PathoNostics B.V. Maastricht, The Nederlands
Simons, Guus;  PathoNostics B.V. Maastricht, The Nederlands
Rijnders, Bart;  Erasmus medical Centrer, Rotterdam > Department of internal medecine > Infectious diseases
Language :
English
Title :
Direct detection of Aspergillus and azole resistance of Aspergillus fumigatus on broncho-alveolar lavage fluid. Validation of a new Aspergillus real-time PCR
Alternative titles :
[fr] Détection directe d'Aspergillus species et de la résistance d'A. fumigatus aux azolés. Validation d'une nouvelle PCR Aspergillus en temps réel
Publication date :
07 January 2015
Journal title :
Journal of Clinical Microbiology
ISSN :
0095-1137
eISSN :
1098-660X
Publisher :
American Society for Microbiology (ASM), Washington, United States - District of Columbia
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
AgentSchap NL [NL]
Available on ORBi :
since 12 March 2015

Statistics


Number of views
102 (7 by ULiège)
Number of downloads
368 (1 by ULiège)

Scopus citations®
 
128
Scopus citations®
without self-citations
121
OpenCitations
 
112

Bibliography


Similar publications



Contact ORBi