Rolling circle amplification and graphene-based sensor-on-a-chip for sensitive detection of serum circulating miRNAs
| Autor: | Tararaj Dharakul, Preedee Pinpradup, Krissana Khoothiam, Deanpen Japrung, Patraporn Luksirikul, Kiatnida Treerattrakoon, Tawin Iempridee, Thanakorn Jiemsakul, Chookiat Tansarawiput |
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| Rok vydání: | 2018 |
| Předmět: |
Circulating mirnas
Materials science Population Microfluidics Biophysics DNA Single-Stranded Computational biology 01 natural sciences Biochemistry Proof of Concept Study law.invention 03 medical and health sciences law Neoplasms Humans Tuberculosis Multiplex education Molecular Biology 030304 developmental biology Oligonucleotide Array Sequence Analysis Detection limit 0303 health sciences education.field_of_study Graphene 010401 analytical chemistry Cell Biology Chip 0104 chemical sciences MicroRNAs Spectrometry Fluorescence Rolling circle replication Nucleic Acid Amplification Techniques HeLa Cells |
| Zdroj: | Analytical biochemistry. 577 |
| ISSN: | 1096-0309 |
| Popis: | In this study, we developed a simple multiplex miRNA detection platform based on rolling circle amplification and the fluorescence quenching property of reduced graphene oxide. The detection platform could be applied on a microfluidics chip with a mobile system controller to eliminate contamination and to facilitate potential use in remote areas. As a proof of concept, two fluorescence-labeled ssDNA tags were used for detection of miR-29a and miR-144*, two miRNAs that are highly expressed in the blood circulation of some patients with cancer or tuberculosis. The circular ssDNA probes in this study were designed to have an advantage over padlock probes as they can be prepared in advance. Our multiplex miRNA detection platform exhibited high sensitivity and selectivity, with a limit of detection of 0.05 pmol. In addition, our platform could detect target miRNAs from the total miRNA population extracted from human serum or a cancer cell line. These results indicated that our miRNA sensor has the potential to provide simple and high throughput miRNA analysis for disease diagnosis and prognosis. |
| Databáze: | OpenAIRE |
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