論文

査読有り 国際誌
2018年9月4日

Purification-Free MicroRNA Detection by Using Magnetically Immobilized Nanopores on Liposome Membrane.

Analytical chemistry
  • Satoshi Fujii
  • ,
  • Koki Kamiya
  • ,
  • Toshihisa Osaki
  • ,
  • Nobuo Misawa
  • ,
  • Masatoshi Hayakawa
  • ,
  • Shoji Takeuchi

90
17
開始ページ
10217
終了ページ
10222
記述言語
英語
掲載種別
DOI
10.1021/acs.analchem.8b01443

MicroRNAs have critical roles in a number of serious diseases and, as a result, are of major interest as clinical diagnostic targets. Conventionally, microRNAs are collected from blood and urine samples and are measured by either quantitative reverse-transcription polymerase chain reaction or microarray. Recently, nanopore sensing techniques have been applied for measuring microRNAs at the single-molecule level. However, existing techniques are technically complex, needing several tools and requiring purification and/or labeling of microRNA samples prior to use. Here we report a method for microRNA detection in a simple procedure requiring neither purification nor labeling. This system utilizes magnetic beads anchored with DNA and nanopores on a liposome membrane. In the presence of the target microRNA, it forms a duplex with complementary DNA, which is then cleaved by a duplex-specific nuclease (DSN). The cleaved DNA, which harbors a liposome on its terminus, is subsequently released from the magnetic bead, fuses to the lipid bilayer on chip, and emits an electrical signal derived from the formation of a nanopore. Because of a property of the DSN, the signals derived from microRNAs are expected to be amplified in an isothermal reaction. Our system possesses the specificity to detect target microRNAs from mixtures containing >106 different microRNA sequences and readily uses blood or urine samples. Although the limit of detection is above 10 nM and needs to be improved for practical diagnosis, because purification and labeling are not required, the presented system proposes a possible schematic for the development of easy and on-site diagnosis.

Web of Science ® 被引用回数 : 4

リンク情報
DOI
https://doi.org/10.1021/acs.analchem.8b01443
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/30091903
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000444060600021&DestApp=WOS_CPL