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Multiplexed Detection of Attomoles of Nucleic Acids Using Fluorescent Nanoparticle Counting Platform
journal contribution
posted on 2017-12-11, 00:00 authored by Xiaojing Pei, Haoyan Yin, Tiancheng Lai, Junlong Zhang, Feng Liu, Xiao Xu, Na LiThe sensitive multiplexed detection
of nucleic acids in a single
sample by a simple manner is of pivotal importance for the diagnosis
and therapy of human diseases.
Herein, we constructed an automatic fluorescent nanoparticle (FNP)
counting platform with a common fluorescence microscopic imaging setup
for nonamplification multiplexed detection of attomoles of nucleic
acids. Taking the advantages of the highly bright, multicolor emitting
FNPs and magnetic separation, the platform enables sensitive multiplexed
detection without the need for extra fluorescent labels. Quantification
for multiplex DNAs, multiplex microRNAs (miRNA), as well as a DNA
and miRNA mixture was achieved with a similar dynamic range, a limit
of detection down to 5 amol (5 μL detection volume), and a 81–115%
spike recovery from different biological sample matrices. In particular,
the sensitivity for multiplex miRNA is by far among the highest without
using amplification or the lock nucleic acid hybridization enhancement
strategy. Results regarding miRNA-141 from four different cell lines
were agreeable with those of the quantitative reverse transcription
polymerase chain reaction. Simultaneous detection of miRNA-141 and
miRNA-21 in four different cell lines yielded consistent results with
publications, indicating the potential for monitoring multiplex miRNA
expression associated with the collaborative regulation of important
cellular events. This work expands the rule set of multiplex nucleic
acid detection strategies and shows promising potential application
in clinical diagnosis.