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A SIMPLE POLYMERASE CHAIN REACTION/RESTRICTION FRAGMENT LENGTH POLYMORPHISM ASSAY CAPABLE OF IDENTIFYING MEDICALLY RELEVANT FILAMENTOUS FUNGI
Citation:
DEAN, T. R., M. J. KOHAN, D. BETANCOURT, AND M. Y. MENETREZ. A SIMPLE POLYMERASE CHAIN REACTION/RESTRICTION FRAGMENT LENGTH POLYMORPHISM ASSAY CAPABLE OF IDENTIFYING MEDICALLY RELEVANT FILAMENTOUS FUNGI. MOLECULAR BIOTECHNOLOGY. Humana Press Incorporated, Clifton, NJ, 31(1):21-28, (2005).
Description:
The accumulating evidence that suggests that numerous unhealthy conditions in the indoor environment are the result of abnormal growth of the filamentous fungi (mold) in and on buildign surfaces. In order to accurately reflect the organisms responsible for these maladies it is of utmost importance to identify them in an accurate and timely manner. To this end, we performed a simple polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) analysis on multiple members of species known to negatively influence the indoor environment. The general analyzed were Stachybotrys, Penicillium, Aspergillus, and Cladosporium. Each organisim underwent PCR with universal primers that amplified ribosomal sequences followed by enzymatic digestion with EcoRI, HaeIII, MspI, and Hinfl. Our results show that using this combination of restriction enzymes enables toe identification of these fungal organisms at the species level. The method is cost effective, easy to perform, and accurate.