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

Mingcheng Xu, Jiawei Ye, Dan Yang, Abdu Ahmed Abdullah AL-maskri, Haihong Hu, Cheulhee Jung, S. Cai, Su Zeng

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

A novel microRNA (miRNA) quantification method has been developed using one-step rolling circle-quantitative 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).

Original languageEnglish
Pages (from-to)208-215
Number of pages8
JournalAnalytica Chimica Acta
Volume1077
DOIs
Publication statusPublished - 2019 Oct 24

Keywords

  • MiR-200a
  • MiRNA detection
  • One-step amplification
  • Rolling circle amplification
  • Rolling circle-quantitative PCR (RC-qPCR)
  • Vent (exo-) DNA polymerase

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy

Fingerprint Dive into the research topics of 'Ultrasensitive detection of miRNA via one-step rolling circle-quantitative PCR (RC-qPCR)'. Together they form a unique fingerprint.

Cite this