Real-time and semiquantitative RT-PCR methods to analyze gene expression patterns during Aspergillus-host interactions.
Aspergillus species are infamous for causing several plant and animal diseases that directly (e.g., invasive aspergillosis) or indirectly (e.g., consumption of toxic food supplies) can lead to high rates of morbidity in humans and animals worldwide. Despite progress in molecular information and manipulation of Aspergillus spp., including genome sequence availability and suitable transformation methodologies, efforts to control Aspergillus diseases are still far from satisfactory, due in part to lack of knowledge of fungal virulence attributes. In order to obtain meaningful insights on the disease mechanism(s), it is essential to detect virulence gene expression during host invasion. Here, we describe two PCR-based detection methods of Aspergillus gene expression in both plant and mammalian tissues. Moreover, these techniques can be employed for routine screening of large numbers of aspergilli to improve diagnosis, disease monitoring, and therapy of fungal disease.
Bok
Jin Woo
JW
Department of Medical Microbiology and Immunology, University of Wisconsin, Madison, Wisconsin, USA.
Keller
Nancy P
NP
Tsitsigiannis
Dimitrios I
DI
D000818:Animals; D010367:Arachis / Q000382:microbiology; D001228:Aspergillosis / Q000382:microbiology; D001230:Aspergillus / Q000235:genetics* / Q000472:pathogenicity*; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D054884:Host-Pathogen Interactions / Q000235:genetics*; D008168:Lung / Q000382:microbiology; D008172:Lung Diseases, Fungal / Q000382:microbiology; D051379:Mice; D008813:Mice, Inbred ICR; D009172:Mycology / Q000379:methods; D012331:RNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D012331:RNA, Fungal