Highly specific DNA detection employing ligation on suspension bead array readout.

Mezger A, K├╝hnemund M, Nilsson M, Herthnek D

N Biotechnol 32 (5) 504-510 [2015-09-25; online 2015-02-11]

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.

Affiliated researcher

PubMed 25681158

DOI 10.1016/j.nbt.2015.01.011

Crossref 10.1016/j.nbt.2015.01.011

pii: S1871-6784(15)00015-1

Publications 7.2.9