Open AccessLabel-Free Isothermal Amplification Assay for Specific and Highly Sensitive Colorimetric miRNA Detection
Author(s) -
Stefano Persano,
María L. Guevara,
Joy Wolfram,
Elvin Blanco,
Haifa Shen,
Mauro Ferrari,
Pier Paolo Pompa
Publication year - 2016
Publication title -
acs omega
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.779
H-Index - 40
ISSN - 2470-1343
DOI - 10.1021/acsomega.6b00109
Subject(s) - loop mediated isothermal amplification , microbiology and biotechnology , breast cancer , naked eye , microrna , chemistry , colloidal gold , cancer cell lines , multiple displacement amplification , cancer , biology , cancer cell , computational biology , dna , chromatography , nanoparticle , detection limit , polymerase chain reaction , gene , biochemistry , nanotechnology , materials science , genetics , dna extraction
We describe a new method for the detection of miRNA in biological samples. This technology is based on the isothermal nicking enzyme amplification reaction and subsequent hybridization of the amplification product with gold nanoparticles and magnetic microparticles (barcode system) to achieve naked-eye colorimetric detection. This platform was used to detect a specific miRNA (miRNA-10b) associated with breast cancer, and attomolar sensitivity was demonstrated. The assay was validated in cell culture lysates from breast cancer cells and in serum from a mouse model of breast cancer.