APPLICATION OF THE SAS ADX INTERFACE FOR OPTIMIZATION OF SINGLE AND MULTIPLEX PCR FOR REAL-TIME PCR DETECTION OF LISTERIA

Authors: Bayles, Darrell

Submitted to: United States Japan Natural Resources Protein Panel
Publication Type: Proceedings
Publication Acceptance Date: 11/30/2002
Publication Date: 6/8/2003
Citation: Bayles, D.O. 2003. Application of the sas adx interface for optimization of single and multiplex pcr for real-time pcr detection of listeria. United States Japan Natural Resources Protein Panel 31st Annual Meeting Proceedings. p. H1-9.

Interpretive Summary:

Technical Abstract: Real time PCR (RT-PCR) offers several advantages over detection methods that rely on separate PCR and gel electrophoresis steps. The SAS Institute ADX interface was used to plan and generate response surface models for the optimization of a multiplex PCR specific for the genus Listeria and for the species L. monocytogenes. The modeled response surfaces allowed the use of interactive SAS tools to evaluate and optimize the PCR reaction conditions while minimizing the number of experimental runs, time, and expense. Following optimization of the multiplex PCR, the system was validated for RT-PCR by assays containing fluorescent probes within the PCR reaction mixture.