Rapid and Sensitive Detection of Repetitive Nucleic Acid Sequences Using Magnetically Modulated Biosensors

Michael Margulis, Amos Danielli

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

Repetitive DNA sequences are abundant in the genome of most biological species. These sequences are naturally "preamplified", which makes them a preferential target for a variety of biological assays. Current methods to detect specific DNA sequences are based on the quantitative polymerase chain reaction (PCR), which relies on target amplification by Taq polymerase and uses a fluorescent resonance energy transfer (FRET)-based probe. Here, to rapidly detect a repetitive DNA sequence, we combine a highly sensitive magnetic modulation biosensing (MMB) system and a modified double-quenched FRET-based probe. The high numbers of copies of the female-specific XhoI sequence of the domestic chicken (Gallus gallus), combined with the low background fluorescence levels of the modified double-quenched probe and the high sensitivity of the MMB system, allow us to determine the chick sex in ovo within 13 min, with 100% sensitivity and specificity. Compared to quantitative PCR, the presented assay is 4-9 times faster. More broadly, by specifically tailoring the primers and probe, the proposed assay can detect any target DNA sequence, either repetitive or nonrepetitive.

Original languageEnglish
Pages (from-to)11749-11755
Number of pages7
JournalACS Omega
Volume4
Issue number7
DOIs
StatePublished - 31 Jul 2019

Bibliographical note

Publisher Copyright:
Copyright © 2019 American Chemical Society.

Funding

The authors thank Shira Roth and Yehudit Michelson for their valuable discussions, and Meir Cohen for technical assistance. This research was supported by the Israel Science Foundation (Grant No. 1142/15) and the Israel Innovation Authority (Kamin, Grant No. 59042). James Ballard provided an editorial review of the manuscript.

FundersFunder number
Israel Innovation Authority59042
Israel Science Foundation1142/15

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