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P2069
Optimizing Nucleic Acid Detection by Altering Detection Chemistry
Presenter Heather Harding, Radix BioSolutions, Ltd, USA
Additional Authors: Bruce Amsden, Kerry Oliver, and Jo Wegstein
Standard amplified nucleic acid hybridization assays require significant in silico and experimental testing to determine the optimal primers and probes for the detection of specific nucleic acid sequences.  However, very little analysis or effort is spent optimizing the detection molecules used for a particular hybridization assay.  Previously, we have demonstrated that the choice of specific phycoerythrin-streptavidin  (PE-StAv) conjugates, used for the detection of nucleic acid hybridization assay, developed for use on the Luminex® xMAP® technology can increase assay sensitivity greatly.  In the present study, we have examined the effects of changing the length of spacer between the xMAP microsphere surface and the hybridizing probe, changing the length of spacer between the amplification primer sequence and the 5’-biotin moiety used to capture the PE-StAv conjugate, and varying the PE-StAv conjugate structure.  The combined effects of these changes demonstrate that varying spacer length and detection conjugate structure enhances greatly the sensitivity of standard nucleic acid hybridization assays performed on xMAP technology.