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DETECTION OF STACHYBOTRYS CHARTARUM USING rRNA, tri5, AND Β-TUBULIN PRIMERS AND DETERMINING THEIR RELATIVE COPY NUMBER BY REAL TIME PCR
Citation:
BLACK, J. A., T. R. DEAN, K. FOARDE, AND M. Y. MENETREZ. DETECTION OF STACHYBOTRYS CHARTARUM USING rRNA, tri5, AND Β-TUBULIN PRIMERS AND DETERMINING THEIR RELATIVE COPY NUMBER BY REAL TIME PCR. Mycological Research. Elsevier BV, AMSTERDAM, Netherlands, 112(7):845-851, (2008).
Impact/Purpose:
Journal article
Description:
This research utilizes the quantitative polymerase chain reaction (qPCR) to determine ribosomal copy number of fungal organisms found in unhealthy indoor environments. Knowing specific copy numbers will allow for greater accuracy in quantification when utilizing current pQCR techniques. The researcher needs to evaluate as many sequences as available for designing species specific PCR primers. This research aligned 11, 9 and 16 DNA sequences entered for Stachybotrys spp. rRNA, tri5 and 16 beta-tubulin regions, respectively. It was possible to align and determine consensus primer sets for the 9 tri5 and the 16 beta-tubulin sequences, but there was no consensus sequence that could be derived from the alignment of the 11 rRNA sequences. However, by judicious clustering of the sequences that aligned well, it was possible to design three sets of primers for the rRNA region of S. chartarum. The two primer sets for tri5 and beta-tubulin produced satisfactory PCR results for all four strains of S. chartarum used in this study while only one rRNA primer set of three produced similar satisfactory results. The rRNA number is approximately 2-log greater than for tri5 and beta-tubulin in the four strains of S. chartarum tested.
