Ultrasensitive detection of miRNA via one-step rolling circle-quantitative PCR (RC-qPCR)

Xu, Mingcheng; Ye, Jiawei; Yang, Dan; AL-Maskri, Abdu Ahmed Abdullah; Hu, Haihong; Jung, Cheulhee; Cai, Sheng; Zeng, Su

doi:10.1016/j.aca.2019.05.028

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Ultrasensitive detection of miRNA via one-step rolling circle-quantitative PCR (RC-qPCR)

Xu, Mingcheng;
Ye, Jiawei;
Yang, Dan;
AL-Maskri, Abdu Ahmed Abdullah;
Hu, Haihong;
... Jung, Cheulhee;
외 2명

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초록

A novel microRNA (miRNA) quantification method has been developed using one-step rolling circlequantitative PCR (RC-qPCR) analysis. Vent (exo-) DNA polymerase is firstly utilized to combine a rolling circle amplification (RCA) and qPCR in one step with high sensitivity and specificity in our RC-qPCR assay. Before performing the RC-qPCR, a padlock probe is ligated only when it is perfectly hybridized with miRNA. This ligation-based miRNA assay is highly specific for mature miRNAs, discriminating among related miRNAs that differ by as little as one nucleotide. It exhibits a dynamic range of seven orders of magnitude with a detection limit of 500 aM, and could be also used for the quantification of other small RNA molecules such as short interfering RNAs (siRNAs). (C) 2019 Elsevier B.V. All rights reserved.

키워드

Rolling circle-quantitative PCR (RC-qPCR); Rolling circle amplification; Vent (exo-) DNA polymerase; One-step amplification; MiR-200a; MiRNA detection; FREE ELECTROCHEMICAL DETECTION; HIGHLY SENSITIVE DETECTION; MESSENGER-RNA; SIGNAL AMPLIFICATION; MICRORNA EXPRESSION; GENES; ASSAY; IDENTIFICATION; MICROCHIP; NANOCUBES

제목: Ultrasensitive detection of miRNA via one-step rolling circle-quantitative PCR (RC-qPCR)

저자: Xu, Mingcheng; Ye, Jiawei; Yang, Dan; AL-Maskri, Abdu Ahmed Abdullah; Hu, Haihong; Jung, Cheulhee; Cai, Sheng; Zeng, Su

DOI: 10.1016/j.aca.2019.05.028

발행일: 2019-10-24

유형: Article

저널명: Analytica Chimica Acta

권: 1077

페이지: 208 ~ 215