Detection of Aspergillus fumigatus by quantitative polymerase chain reaction in air samples impacted on low-melt agar

Background The standard procedure for routine environmental sampling for the prevention of invasive aspergillosis outbreaks is culturing of Aspergillus fumigatus after impaction of air. Time to results is usually 7 days. A preliminary study was carried out to compare the time to results and sensitiv...

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Bibliographic Details
Published in:American journal of infection control 2010-04, Vol.38 (3), p.195-198
Main Authors: Bellanger, Anne-Pauline, PharmD, Reboux, Gabriel, PhD, Murat, Jean-Benjamin, PharmD, Bex, Valerie, PhD, Millon, Laurence, PharmD, PhD
Format: Article
Language:English
Subjects:
Agar
Air Microbiology
Airborne particulates
Algae
Aspergillus fumigatus
Aspergillus fumigatus - isolation & purification
Bacteriological Techniques - methods
Biological and medical sciences
Culture Media - chemistry
Disease control
environmental sampling
Epidemics
Epidemiology. Vaccinations
fungal surveillance
Fungi
General aspects
Humans
immunocompromised patients
Immunodeficiencies
Immunodeficiencies. Immunoglobulinopathies
Immunopathology
Infection Control
Infectious Disease
Infectious diseases
Life Sciences
Medical sciences
Microbiology and Parasitology
Mycology
Polymerase chain reaction
Polymerase Chain Reaction - methods
QPCR
Sensitivity and Specificity
Studies
Time Factors
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Summary:Background The standard procedure for routine environmental sampling for the prevention of invasive aspergillosis outbreaks is culturing of Aspergillus fumigatus after impaction of air. Time to results is usually 7 days. A preliminary study was carried out to compare the time to results and sensitivity of culturing and quantitative polymerase chain reaction (QPCR) in the detection of airborne A fumigatus. Methods Fungal DNA was extracted from 43 samples of impacted low-melt agar by a 3-step extraction method and amplified by QPCR. Identification was made using a specific A fumigatus probe. Results With QPCR, 19 of the 43 samples were positive for A fumigatus ; with culturing, 7 of these 19 samples were positive, and 12 were negative. The cycle threshold (Ct) values for the 12 culture-negative samples were between 39 and 43 cycles, and the Ct values for 6 of the 7 culture-positive samples were
ISSN:0196-6553
1527-3296
DOI:10.1016/j.ajic.2009.08.003