MicroRNA detection using lateral flow nucleic acid strips with gold nanoparticles

• Published in: Talanta (Oxford) 2012-09, Vol.99, p.375-379
• Main Authors: Hou, Shao-Yi, Hsiao, Yi-Ling, Lin, Ming-Shu, Yen, Chun-Che, Chang, Chi-Sheng
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Base Sequence; Biocatalysis; Biosensing Techniques - methods; Chemistry; Exact sciences and technology; Gold - chemistry; Gold nanoparticles; Lateral flow nucleic acid strips; Limit of Detection; Metal Nanoparticles - chemistry; MicroRNA detection; MicroRNAs - analysis; MicroRNAs - chemistry; Mung-bean nuclease; Nucleic Acid Hybridization; Oligonucleotide Probes - chemistry; Oligonucleotide Probes - genetics; Plant Proteins - metabolism; Point-of-Care Systems; Single-Strand Specific DNA and RNA Endonucleases - metabolism; Single-strand-specific nuclease; Sulfhydryl Compounds - chemistry; Time Factors
• ISSN: 0039-9140; 1873-3573
• DOI: 10.1016/j.talanta.2012.05.067

In this study, the tested microRNA and the detection probe perfectly match with the capture probe instead of the traditional sandwich methods in which the tested oligonucleotide matches with the detection and capture probes. To avoid non-specific signals, mung-bean nuclease, a single-strand-specific nuclease, catalyzes the degradation of the capture probe if there is no tested miRNA in the samples. The gold nanoparticles conjugate the thiol-DNA as the detection probe and the biotin-single strand DNA serves as the capture probe. The avidin–biotin–Au-sample complex is captured by the anti-avidin antibody immobilized on a flow strip. The detection and quantification of the gold nanoparticle signal indicate the existence and quantity of the target miRNA. One fmol and five amol of the synthetic microRNA were detected without and with the silver enhancement, respectively. This highly sensitive and specific assay takes about 70min after the RNA purification and preparation. It is simple, convenient, fast, and suitable for point-of-care. ► In this study, a fast and sensitive method is developed to detect microRNA. ► One fmol of the tested microRNA were detected without the silver enhancement. ► Five amol of the tested microRNA were detected with the silver enhancement. ► This method does not need equipments. ► It is simple, convenient, fast, and suitable for point-of-care.