Direct detection of circulating microRNAs in serum of cancer patients by coupling protein-facilitated specific enrichment and rolling circle amplification.

We have developed a simple method for direct detection of circulating microRNAs in serum by using the p19 protein-functionalized magnetic beads (PFMBs) for specific enrichment and rolling circle amplification (RCA). The detection limit for microRNAs is 1 fM. Therefore, the proposed method has the potential of being used in the analysis of circulating microRNAs and clinical diagnosis.

[1]  Feng-jun Wang,et al.  Correlation and quantitation of microRNA aberrant expression in tissues and sera from patients with breast tumor. , 2010, Gynecologic oncology.

[2]  C. Croce,et al.  Frequent deletions and down-regulation of micro- RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[3]  Chun-yang Zhang,et al.  Sensitive detection of microRNAs with hairpin probe-based circular exponential amplification assay. , 2012, Analytical chemistry.

[4]  Yongqiang Cheng,et al.  Ultrasensitive detection of microRNAs by exponential isothermal amplification. , 2010, Angewandte Chemie.

[5]  Qian Wang,et al.  High specific and ultrasensitive isothermal detection of microRNA by padlock probe-based exponential rolling circle amplification. , 2013, Analytical chemistry.

[6]  M. Berezovski,et al.  Three-mode electrochemical sensing of ultralow microRNA levels. , 2012, Journal of the American Chemical Society.

[7]  Jing Zhang,et al.  Highly sensitive multiple microRNA detection based on fluorescence quenching of graphene oxide and isothermal strand-displacement polymerase reaction. , 2012, Analytical chemistry.

[8]  Yu-Qiang Liu,et al.  One-step, multiplexed fluorescence detection of microRNAs based on duplex-specific nuclease signal amplification. , 2012, Journal of the American Chemical Society.

[9]  Chunhai Fan,et al.  DNAzyme-based rolling-circle amplification DNA machine for ultrasensitive analysis of microRNA in Drosophila larva. , 2012, Analytical chemistry.

[10]  F. Slack,et al.  Oncomirs — microRNAs with a role in cancer , 2006, Nature Reviews Cancer.

[11]  Zhengping Li,et al.  Highly sensitive determination of microRNA using target-primed and branched rolling-circle amplification. , 2009, Angewandte Chemie.

[12]  Zhiqiang Gao,et al.  A label-free biosensor for electrochemical detection of femtomolar microRNAs. , 2013, Analytical chemistry.

[13]  C. Burge,et al.  Most mammalian mRNAs are conserved targets of microRNAs. , 2008, Genome research.

[14]  A. Mulchandani,et al.  Electronic detection of microRNA at attomolar level with high specificity. , 2013, Analytical chemistry.

[15]  X Chris Le,et al.  DNA-mediated homogeneous binding assays for nucleic acids and proteins. , 2013, Chemical reviews.

[16]  X. Chen,et al.  Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases , 2008, Cell Research.

[17]  K. Livak,et al.  Real-time quantification of microRNAs by stem–loop RT–PCR , 2005, Nucleic acids research.

[18]  Nóra Varga,et al.  Sensitive and specific detection of microRNAs by northern blot analysis using LNA-modified oligonucleotide probes. , 2004, Nucleic acids research.

[19]  C. Perou,et al.  A custom microarray platform for analysis of microRNA gene expression , 2004, Nature Methods.

[20]  G. Szittya,et al.  Size Selective Recognition of siRNA by an RNA Silencing Suppressor , 2003, Cell.

[21]  D. Bartel MicroRNAs: Target Recognition and Regulatory Functions , 2009, Cell.

[22]  M. Berezovski,et al.  Quantitative analysis of microRNA in blood serum with protein-facilitated affinity capillary electrophoresis. , 2011, Analytical chemistry.

[23]  Sota Asaga,et al.  Direct serum assay for microRNA-21 concentrations in early and advanced breast cancer. , 2011, Clinical chemistry.

[24]  Cheng Zhang,et al.  Backbone-modified molecular beacons for highly sensitive and selective detection of microRNAs based on duplex specific nuclease signal amplification. , 2013, Chemical communications.