Genetic assay for point-of-care applications should be simple, specific, sensitive, fast, and cost-effective. Assays using DNA-based multicomponent nucleic acid enzymes, MNAzymes, satisfy most of these requirements, but need fairly expensive and complex fluorometer apparatus for readout. Here, we present a novel platform that combines MNAzyme-based detection with absorbance properties of gold nanoparticles to allow for simple colorimetric readout.The assay is performed in solution with 50 pM sensitivity without the need for purification or separation steps, can detect multiple DNA or RNA targets in parallel, and is easily adaptable to new targets. We demonstrate that this technology is capable of rapid and specific detection of synthetic targets for syphilis, malaria, hepatitis B, gonorrhea, and other bacterial infections. Furthermore, we apply the assay to detect a panel of 5 different bacterial pathogens that are responsible for the majority of urinary tract infections (UTI). UTI pathogens spiked into urine are detected in parallel with clinically relevant sensitivity of 104-105 bacteria/mL. All the reagents can be lyophilized for long-term storage and transport, making the assay well suited for point-of-care applications.

Plasmonic DNAzyme strategy for point-of-care detection of genetic signatures from infectious pathogens

247th National Meeting (2014)

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