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Utilizing Pyrosequencing and Quantitative pCR to Characterize Fungal Populations among House Dust Samples
Nonnenmann, M. W., G. Coronado, B. Thompson, W. C. Griffith, J. D. Hanson, S. J. VESPER, AND E. M. Faustman. Utilizing Pyrosequencing and Quantitative pCR to Characterize Fungal Populations among House Dust Samples. JOURNAL OF ENVIRONMENTAL MONITORING. Royal Society of Chemistry, Cambridge, Uk, 14(8):2038-2043, (2012).
Molecular techniques are an alternative to culturing and counting methods in quantifying indoor fungal contamination. Pyrosequencing offers the possibility of identifying unexpected indoor fungi. In this study, 50 house dust samples were collected from homes in the Yakima Valley, WA. Each sample was analyzed by quantitative PCR (QPCR) for 36 common fungi and by fungal tag-encoded flexible (FLX) amplicon pyrosequencing (fTEFAP) for these and additional fungi. Only 24 of the samples yielded amplified results using fTEFAP but QPCR successfully amplified all 50 samples. Over 450 fungal species were detected by fTEFAP but most were rare. Twenty-two fungi were found by fTEFAP to occur with at least an average of ≥0.5% relative occurrence. Many of these fungi seem to be associated with plants, soil or human skin. Combining fTEFAP and QPCR can enhance studies of fungal contamination in homes.
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Record Details:Record Type: DOCUMENT (JOURNAL/PEER REVIEWED JOURNAL)
Organization:U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
NATIONAL EXPOSURE RESEARCH LAB
MICROBIOLOGICAL AND CHEMICAL EXPOSURE ASSESSMENT DIVISION
MICROBIAL EXPOSURE RESEARCH BRANCH