In the present study, we developed a highly sensitive and convenient biosensor consisting of gold nanoparticle （AuNP） probes and a gene chip to detect microRNAs （miRNAs）. Specific oligonucleotides were attached to...In the present study, we developed a highly sensitive and convenient biosensor consisting of gold nanoparticle （AuNP） probes and a gene chip to detect microRNAs （miRNAs）. Specific oligonucleotides were attached to the glass surface as capture probes for the target miRNAs, which were then detected via hybridization to the AuNP probes. The signal was amplified via the re- duction of HAuCI4 by H202. The use of a single AuNP probe detected 10 pmol L-1 of target miRNA. The recovery rate for miR-126 from fetal bovine serum was 81.5%-109.1%. The biosensor detection of miR-126 in total RNA extracted from lung cancer tissues was consistent with the quantitative PCR （qPCR） results. The use of two AuNP probes further improved the de- tection sensitivity such that even 1 fmol L-t of target miR-125a-5p was detectable. This assay takes less than 1 h to complete and the results can be observed by the naked eye, The platform simultaneously detected lung cancer related miR-126 and miR-125a-5p. Therefore, this low cost, rapid, and convenient technology could be used for ultrasensitive and robust visual miRNA detection.展开更多
基金supported by the National Basic Research Program of China (2012CB933303)the National Natural Science Foundation of China (61571429, 61571077, 61401442)+2 种基金the Innovation Team of Henan University of Science and Technology (2015XTD003)the Science and Technology Commission of Shanghai Municipality (12441902600, 1402H233900)the Shanghai Clinical Center/Shanghai Xuhui Central Hospital, Chinese Academic of Sciences (BRC2012002)
文摘In the present study, we developed a highly sensitive and convenient biosensor consisting of gold nanoparticle （AuNP） probes and a gene chip to detect microRNAs （miRNAs）. Specific oligonucleotides were attached to the glass surface as capture probes for the target miRNAs, which were then detected via hybridization to the AuNP probes. The signal was amplified via the re- duction of HAuCI4 by H202. The use of a single AuNP probe detected 10 pmol L-1 of target miRNA. The recovery rate for miR-126 from fetal bovine serum was 81.5%-109.1%. The biosensor detection of miR-126 in total RNA extracted from lung cancer tissues was consistent with the quantitative PCR （qPCR） results. The use of two AuNP probes further improved the de- tection sensitivity such that even 1 fmol L-t of target miR-125a-5p was detectable. This assay takes less than 1 h to complete and the results can be observed by the naked eye, The platform simultaneously detected lung cancer related miR-126 and miR-125a-5p. Therefore, this low cost, rapid, and convenient technology could be used for ultrasensitive and robust visual miRNA detection.
