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Inhibition of quantitative PCR analysis of fungal conidia associated with indoor air particulate matter.

Authors
McDevitt-JJ; Lees-PSJ; Merz-WG; Schwab-KJ
Source
Aerobiologia 2007 Jan; 23(1):35-45
NIOSHTIC No.
20037416
Abstract
Environmental samples analyzed by quantitative PCR (qPCR) are subject to interference by inhibitors present in the environment being sampled. A controlled determination of the effect of inhibitors associated with sampling indoor air and the ability of internal standard controls to detect inhibition was carried out by filter collection of air samples followed by spiking of the filters with green fluorescent protein-expressing Aspergillus fumigatus conidia. Microscopic conidial counts were compared with qPCR results and correlated with levels of particulate matter and viable airborne microorganisms. Our data showed that PCR can be inhibited by masses of particulate matter as low as 50 g and that the amount of inhibition was positively correlated with the mass of particulate (r = 0.75) and the number of non-filamentous organisms (r = 0.73). The use of internal standard DNA identified the presence of inhibitors and indicated the need for additional sample processing or qualification of sample results.
Keywords
Fungi; Environmental-factors; Air-samples; Airborne-particles; Analytical-processes; Analytical-methods; Particulates; Particulate-sampling-methods; Sampling; Sampling-methods; Microorganisms; Author Keywords: Airborne fungi; Aspergillus fumigatus; Conidia; Quantitative PCR; Particulate-matter; Sample inhibition
Contact
K. J. Schwab, Department of Environmental Health Sciences, The Johns Hopkins University, Bloomberg School of Public Health, 615 N. Wolfe St., Baltimore, MD 21205, USA
CODEN
AROBFT
Publication Date
20070101
Document Type
Journal Article
Email Address
kschwab@jhsph.edu
Funding Type
Grant
Fiscal Year
2007
NTIS Accession No.
NTIS Price
Identifying No.
Grant-Number-T42-OH-008428
Issue of Publication
1
ISSN
0393-5965
Source Name
Aerobiologia
State
MA; MD
Performing Organization
Johns Hopkins University
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