High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR

Goodman, Laura B.; Anderson, Renee R.; Slater, Marcia; Ortenberg, Elen; Renshaw, Randall W.; Chilson, Brittany D.; Laverack, Melissa A.; Beeby, John S.; Dubovi, Edward J.; Glaser, Amy L.

This procedure has been used for routine testing of respiratory pathogens in our laboratory over the course of six months (2-3 times per week). We have also had success using the same procedure for enteric pathogen profiling in fecal samples and bacterial isolates on a separately customized plate. Once the plates are produced, experienced staff can complete steps 2-7 within one standard workday. The most critical steps are the proper sealing of the preamp plate, the transfer of preamplified samples between plates (which must be done rapidly to prevent evaporation), and the final covering of the amplification plate. Use of the macro spreadsheet as a guide for result analysis (checking curves for samples and controls) was also critical as it greatly reduced the amount of time and paperwork needed for this process. The provided macro spreadsheet is an example; it would need to be modified or re-created for plates with different configurations. This can easily be performed by someone with basic spreadsheet knowledge.