This research attempts to develop a molecular bioassay that uses a Polymerase Chain Reaction (PCR) technique for detecting fungi commonly found in an indoor environment. During sample preparation, the method of bead-beating was identified as the most effective way for spore breakage and fungal DNA release. In the process of assay development, a fungal primer set, FF2/FRl, was designed and tested with DNA from human, rats, mice, bacteria, pollens and six fungal species. The results based on the crude extracts indicated that the primer set successfully amplified the fungal DNA without any cross-amplification with non-fungal DNA. In addition, higher amplification efficiencies were achieved using 20% nutrient media, rather than water, as the process solution. This PCR assay has a sensitivity of detecting low levels of fungi (2 fungal spores per reaction).
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