The ability to detect single nucleotide polymorphisms (SNPs) in DNA and RNA sequences has broad implications in genomic studies and disease diagnosis. A convenient detection method for specific nucleic acid sequences, therefore, is important in the detection of bacteria and viruses, SNPs leading to genetically defined outcomes, and in the development of increasingly patient-centric pharmaceuticals. Current methods for detection of these sequences are expensive, as well as labor intensive and time consuming. An ideal approach provides a straightforward, inexpensive, and disposable detection format. The technology suite described here provides methodologies for room temperature detection of specific nucleic acid sequences via a modular, label-free probe. The probe changes the assay solution’s color only in the presence of the exact, targeted DNA sequence. As such, the technology suite provides a low cost, easy to use genetic variant detection platform that is quick (the reaction takes less than 30 minutes), can be done at room temperature, and can detect single nucleotide differences. Further, these methodologies may also allow for improved pharmaceutical development and the early detection of bacteria, viruses, and cancer.
SNPs and RNA/DNA fragments provide a highly specific target for the detection of certain genetic variations and viral and bacterial disease markers. Conventional technologies use expensive fluorescently labeled TaqMan and molecular beacon (MB) probes. Unlike current strategies, this technology can use unlabeled probes which are up to 40 times less expensive to synthesize. Compared to MB-based probes, this technology demonstrates higher hybridization selectivity at ambient temperatures and therefore more accurate disease diagnosis. In one variation, the technology provides a PCR-free fluorescent output that enables the detection of pathogenic bacteria at levels 1000 times lower than MB probes and other hybridization probes. In another variation, the technology allows rapid visualization of the presence of pathogenic bacteria via colorimetric change, making point of care determinations of disease possible via naked eye evaluation.
These technologies have been evaluated in a laboratory setting and have demonstrated highly reproducible and sensitive outcomes.
Dmitry Kolpashchikov, Ph.D.
Patent Issued (US 8,551,678)
Patent Issued (US 8,313,903)
Patent Issued (US 8,354,227)
Tech Ventures Reference: IR 1941, IR 1953, IR1884, IR 2462