Development of a Fungus-Specific PCR Assay for Detecting Low-Level Fungi in an Indoor Environment

Details

• Personal Author:
  Chen B ; Ong T ; Whong, Wen-Zong ; Zhou G
• Description:
  A fungus-specific PCR assay using only one primer set has been developed for detecting indoor fungi. Four fungal primer sets, NS3/NS4, NS5/NS6, FF1/FR1 and FF2/FR1, were tested with DNA from humans, rats, mice, bacteria, pollens and six commonly found fungal species (Alternaria chamydospora, Aspergillus flavus, Candida famata, Cladosporium fermentans, Penicillium chrycoIgenum and Stachybotrys chartarum). Results indicated that, although all four primer sets could amplify the fungal DNA, only FF2/FR1 demonstrated no cross-amplification with non-fungal DNA. In addition, these amplified fragments were sequenced to ensure that they indeed matched known fungal DNA sequences. Furthermore, besides the tested fungi, eighteen more genera of fungal sequences were examined and found to match the FF2/FR1. Here, the method of bead-beating was identified as the most effective way for spore breakage and fungal DNA release. The PCR amplification efficiency and potential inhibition were examined using different process solutions and preparation procedures. It was found that, when using 20% nutrient media and homogenization-first procedure, a higher amplification efficiency with less inhibition was achieved. Although positive bands were observed at 0.2 fungal spore/reaction using the homogenization-first procedure, the sensitivity of this assay would be two fungal spores/reaction for environmental samples. [Description provided by NIOSH]
• Subjects:
  Animals Environment Fungi Fungicide, Industrial Laboratories Microbiology Occupational Health Safety
• Keywords:
  18 S RRNA Animal Studies Author Keywords: Fungus-specific Primer Bacteria Bacterial Cultures Environmental Factors Fungicides Indoor Air Quality Indoor Fungi Laboratory Animals Microorganisms PCR Sampling

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• ISSN:
  0890-8508
• Document Type:
  Text
• Genre:
  Journal Article
• Place as Subject:
  OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  HELD - Health Effects Laboratory Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  339-348
• Volume:
  14
• Issue:
  6
• NIOSHTIC Number:
  nn:20020872
• Citation:
  Mol Cell Probes 2000 Dec; 14(6):339-348
• Contact Point Address:
  Health Effects Laboratory Division, NIOSH, 1095 Willowdale Road, Morgantown, WV 26505, USA
• Email:
  bdc4@cdc.gov
• Federal Fiscal Year:
  2001
• Peer Reviewed:
  True
• Source Full Name:
  Molecular and Cellular Probes
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:d8b9e636120b11c0fd82e943e2d490aa6f2420affa93c00bb646a94870630631f803009a9e9d7c6333bacf23885d202f2dab2e8f867f9f8158e2f28bf98c1fca
• Download URL:
  https://stacks.cdc.gov/view/cdc/198529/cdc_198529_DS1.pdf
• File Type:
  [PDF - 139.39 KB ]