Title: Development and Use of Fluorescent Antibody and qPCR Protocols for the Electrostatic Spore Trap Authors
Submitted to: National American Phytopathology Meetings
Publication Type: Abstract Only
Publication Acceptance Date: July 13, 2009
Publication Date: June 1, 2010
Citation: Schneider, R.W., Haudenshield, J.S., Hartman, G.L., Mahaffee, W.F. 2010. Development and Use of Fluorescent Antibody and qPCR Protocols for the Electrostatic Spore Trap [abstract]. American Phytopathological Society National Meeting, August 1-5, 2009, Portland, Oregon. 99:S115. Technical Abstract: Fluorescent antibody (FA) and qPCR protocols were evaluated for the newly developed aerobiological sampler (Ionic Spore Trap), which depends upon electrostatic deposition of particulates onto a 25 mm aluminum disk (stub). This device was originally designed for assessment of captured particulates by scanning electron microscopy (SEM), which could be augmented with automated scanning of stubs and machine vision. The trap was used to monitor airborne populations of Phakopsora pachyrhizi, causal agent of soybean rust, in Florida and Louisiana in 2008. Urediniospores were observed by SEM analyses several weeks before first symptoms were found at both locations. A standard compound microscope also may be used with clear adhesive tape affixed to the stub. We now report on the successful development of FA and qPCR protocols, which greatly enhance the versatility and utility of this device. FA detections were conducted with clear double-stick tape and with adhesive carbon disks affixed to the stubs. These protocols will be especially useful for fungi in which spore morphology may not be adequate for identification to species or biotype levels, and they allow for more precise quantification. In addition, a SEM and trained microscopist are not needed for these protocols.