Quantification of microRNA by gold nanoparticle probes.
MicroRNAs are important posttranscriptional regulators of gene expression in animals and plants. A sensitive and specific detection method is urgently needed for intensive studies on differential expression and regulatory roles of microRNA. Here we present a simple and reliable method for the quantification of microRNA. The hybridization products of target microRNA with capture probe and gold nanoparticle probe are immobilized onto the surface of a streptavidin-coated microplate, and the signal is amplified by silver enhancement. Distribution of miR-122a/miR-128 in mouse brain and liver tissue was detected by this method, and synthetic miRNA122a was quantified. This method allowed a lower detect limit of 10 fM with a linear dynamic range from 10 pM to 10 fM and a high specificity to discriminate one single oligonucleotide mismatch of the target microRNA.
Yang
Wen-Jie
WJ
MOE Key Laboratory of Environment and Health, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, wuhan 430030, China.
Li
Xiao-Bo
XB
Li
Yuan-Yuan
YY
Zhao
Li-Fan
LF
He
Wen-Lei
WL
Gao
Yu-Qiu
YQ
Wan
Yan-Jian
YJ
Xia
Wei
W
Chen
Tian
T
Zheng
Hao
H
Li
Miao
M
Xu
Shun-Qing
SQ
D000818:Animals; D001921:Brain / Q000378:metabolism; D006046:Gold / Q000737:chemistry*; D008099:Liver / Q000378:metabolism; D053768:Metal Nanoparticles / Q000737:chemistry*; D051379:Mice; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics; D036103:Nanotechnology / Q000379:methods; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction
D035683:MicroRNAs; D006046:Gold