Utilization of the Stratagene

Abstract
Rapid and accurate methods are needed to quickly detect mycotoxigenic
fungi such as those in the genera of Aspergillus, Penicillium, and Fusarium.
This study was conducted to develop and test a multiplex real-time
qPCR assay to detect and quantify these agriculturally and economically
important fungi. PCR primers were designed to amplify the internal
transcribed spacer (ITS) regions of rDNA from three mycotoxigenic
genera. Genus-specific probes were designed from the ITS sequences of
the most important mycotoxigenic species of Fusarium, Penicillium, and
Aspergillus. Fluorophores were attached to the 5’ end of the probes, which
were detectable with the Stratagene Mx3000P QPCR system. The linear
detection range of the multiplex assay was between 10 pg to 10 ng of
DNA. This assay could be used as an initial step to evaluate the potential
for mycotoxin accumulation in various agricultural commodities.