title: Highly specific DNA detection employing ligation on suspension bead array readout.
abstract: We show for the first time that monomerized rolling circle amplification (RCA) products can be directly detected with the Luminex suspension bead array readout without the need of PCR amplification. Furthermore, using monomerized RCA products to guide ligation of the detection oligonucleotide (DO) to barcode sequences on the magnetic Luminex beads, combined with efficient washing and increased measurement temperature, yields a higher signal to noise ratio. As a proof-of-principle, we demonstrate detection of pathogenic DNA sequences with high reproducibility, sensitivity and a dynamic range over four orders of magnitude. Using padlock probes in combination with bead suspension arrays opens up the possibility for highly multiplexed DNA targeting and readout.
authors: Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, Solna, Sweden.
authors: Anja
authors: A
authors: Mezger
authors: Science for Life Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University, Sweden.
authors: Malte
authors: M
authors: Kühnemund
authors: Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, Solna, Sweden.
authors: Mats
authors: M
authors: Nilsson
authors: Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, Solna, Sweden. Electronic address: david.herthnek@scilifelab.se.
authors: David
authors: D
authors: Herthnek
mesh_terms: D004247:DNA / Q000032:analysis*; D008026:Ligation; D057230:Limit of Detection; D015203:Reproducibility of Results
chemical_list: D004247:DNA