A method has been developed for delivering reagents, including primers, using microbeads that bind the reagents until released into solution with the application of heat or chemicals. Multiple, segregated reaction wells are formed on a single aluminum oxide membrane (AOM) that is used to purify DNA from the sample. Target sequence-specific primers anchored to a solid support, such as functionalized paramagnetic microbeads, are used to anchor the primers during storage, sample preparation, and wash steps. Each well is preloaded with beads containing a different, specific set of forward and reverse PCR primers allowing for parallel singleplex reactions on the same chip. Each reaction uses an intercalating dye for detection, reducing the optimization time greatly as compared to traditional multiplex PCR that requires multiple primer/probe sets with different fluorophores.
Related technologies have been developed that increase multiplexing capability by incorporating dye stability as a measurement (16-0015) and utilize non-cylindrical bead well geometries to improve assay performance and readout (15-0131).
• Uses beads to deliver reagents, including primers, in a simple, parallel amplification and detection method
• Can be used for highly sensitive real-time PCR detection of multiple target sequences on a single chip with or without the use of sequence-specific probes
• May be used clinically to evaluate the presence of disease causing organisms, gene expression, genotyping, or forensic science applications
- An automated integrated platform for rapid and sensitive multiplexed protein profiling using human saliva samples Royal Society of Chemistry, Lab on a Chip Lab Chip, 2014, 14, 1087-1098;DOI: 10.1039/C3LC51303C, Paper