NDLI: Evaluation of interference to conventional and real-time PCR for detection and quantification of fungi in dust

Content Provider	Royal Society of Chemistry (RSC)
Author	Keswani, Jyoti Chen, Bean T. Kashon, Michael L.
Copyright Year	2005
Abstract	Advances in polymerase chain reaction (PCR) have permitted accurate, rapid and quantitative identification of microorganisms in pure cultures regardless of viability or culturability. In this study, a simple sample processing method was investigated for rapid identification and quantification of fungal spores from dust samples using both conventional and real-time PCR. The proposed method was evaluated for susceptibility to interference from environmental dust samples. Stachybotrys chartarum and Aspergillus fumigatus were used as test organisms. The sensitivity of detection in pure culture was 0.1 spore DNA equivalents per PCR reaction corresponding to 20 spores ml−1 in the sample. However, 1 spore DNA equivalent per PCR reaction corresponding to 200 spores ml−1 in the sample was the lowest amount of spores tested without interference in dust samples spiked with spores of either fungal species. The extent of inhibition was calculated using conventional and real-time PCR reactions containing fungal spores, specific primers, specific probes (for real-time PCR) and various amounts of dust. The results indicate that the extent of inhibition by dust on PCR varies with the type and amount of dust, and number of spores. No interference in the analysis of spiked samples was detected from 0.2 mg ml−1 of four real-life dust samples at p-value >0.05 using 2 × 104 spores for conventional PCR and 2 × 105 spores for real-time PCR. However, samples containing >0.2 mg ml−1 real-life dust compromised the PCR assay. These results suggest the potential usefulness of a simple sample processing method in conjunction with PCR for monitoring the fungal content of aerosols collected from indoor environments.
Starting Page	311
Ending Page	318
Page Count	8
File Format	HTM / HTML PDF
ISSN	14640325
Volume Number	7
Issue Number	4
Journal	Journal of Environmental Monitoring
DOI	10.1039/b415498c
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Subscribed
Subject Keyword	DNA polymerase Chain reaction Polymerase chain reaction Stachybotrys Aspergillus fumigatus Conidium DNA No Interference P-value
Content Type	Text
Resource Type	Article
Subject	Public Health, Environmental and Occupational Health Management, Monitoring, Policy and Law

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