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

doi:10.1016/j.aca.2019.05.028

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

^*Corresponding author for this work

Department of Biotechnology

Research output: Contribution to journal › Article › peer-review

41 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 language	English
Pages (from-to)	208-215
Number of pages	8
Journal	Analytica Chimica Acta
Volume	1077
DOIs	https://doi.org/10.1016/j.aca.2019.05.028
Publication status	Published - 2019 Oct 24

Bibliographical note

Publisher Copyright:
© 2019 Elsevier B.V.

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
Environmental Chemistry
Biochemistry
Spectroscopy

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10.1016/j.aca.2019.05.028

Cite this

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

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).",

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

author = "Mingcheng Xu and Jiawei Ye and Dan Yang and \{Abdullah AL-maskri\}, \{Abdu Ahmed\} and Haihong Hu and Cheulhee Jung and S. Cai and Su Zeng",

note = "Publisher Copyright: {\textcopyright} 2019 Elsevier B.V.",

year = "2019",

month = oct,

day = "24",

doi = "10.1016/j.aca.2019.05.028",

language = "English",

volume = "1077",

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journal = "Analytica Chimica Acta",

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T1 - Ultrasensitive detection of miRNA via one-step rolling circle-quantitative PCR (RC-qPCR)

AU - Xu, Mingcheng

AU - Ye, Jiawei

AU - Yang, Dan

AU - Abdullah AL-maskri, Abdu Ahmed

AU - Hu, Haihong

AU - Jung, Cheulhee

AU - Cai, S.

AU - Zeng, Su

PY - 2019/10/24

Y1 - 2019/10/24

N2 - 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).

AB - 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).

KW - MiR-200a

KW - MiRNA detection

KW - One-step amplification

KW - Rolling circle amplification

KW - Rolling circle-quantitative PCR (RC-qPCR)

KW - Vent (exo-) DNA polymerase

UR - https://www.scopus.com/pages/publications/85065848277

U2 - 10.1016/j.aca.2019.05.028

DO - 10.1016/j.aca.2019.05.028

M3 - Article

C2 - 31307711

AN - SCOPUS:85065848277

SN - 0003-2670

VL - 1077

SP - 208

EP - 215

JO - Analytica Chimica Acta

JF - Analytica Chimica Acta

ER -

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

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

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