Ultrasensitive detection of lung cancer-associated miRNAs by multiple primer-mediated rolling circle amplification coupled with a graphene oxide fluorescence-based (MPRCA-GO) sensor.
PMID 31123738 · The Analyst · 2019 · DOI 10.1039/c9an00517j
Authors: Krissana Khoothiam; Kiatnida Treerattrakoon; Tawin Iempridee; Patraporn Luksirikul; Tararaj Dharakul; Deanpen Japrung
Abstract
MicroRNAs (miRNAs) play important roles in gene regulation and have been reported as biomarkers in cancer diagnosis. Herein, we develop an isothermal miRNA detection platform based on the highly efficient, multiple primer-mediated rolling circle amplification method coupled with a graphene oxide-based fluorescence (MPRCA-GO) assay, using lung cancer-associated miRNAs (miR-21 and miR-210) and a reference miRNA (miR-16) as model targets. The combination of the designed ssDNA probe and T4 RNA ligase (T4 Rnl2) used in the MPRCA-GO assay allowed for single-base mismatch discrimination. In addition, the superfluorescence quenching ability of GO allowed for rapid fluorescence detection. The developed platform had a limit of detection as low as 0.87 fM and could detect target miRNAs in cancer cell lines and human serums. Therefore, the MPRCA-GO sensor has the potential for single nucleotide polymorphism (SNP) analysis and applications in clinical diagnostics.
Bacteriophage T4; Biomarkers, Tumor; Cell Line, Tumor; DNA Probes; DNA, Single-Stranded; Fluorescence; Graphite; Humans; Limit of Detection; MicroRNAs; Nucleic Acid Amplification Techniques; Nucleic Acid Hybridization; RNA Ligase (ATP); Spectrometry, Fluorescence; Viral Proteins
Cite
Krissana Khoothiam; Kiatnida Treerattrakoon; Tawin Iempridee; Patraporn Luksirikul; Tararaj Dharakul; Deanpen Japrung. Ultrasensitive detection of lung cancer-associated miRNAs by multiple primer-mediated rolling circle amplification coupled with a graphene oxide fluorescence-based (MPRCA-GO) sensor. The Analyst (2019). PMID 31123738. DOI 10.1039/c9an00517j.