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

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

36 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

Funding Information:
We acknowledge financial support from the National Natural Science Foundation of China (Grant 81573389 ), the National Key R&D Program of China ( 2017YFC0908600 ), the Zhejiang Provincial Natural Science Foundation of China ( LY18H300003 ). Scientific Research Fund of Zhejiang Provincial Education Department ( Y201430444 ).

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
Biochemistry
Environmental Chemistry
Spectroscopy

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10.1016/j.aca.2019.05.028

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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).",

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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 = "Funding Information: We acknowledge financial support from the National Natural Science Foundation of China (Grant 81573389 ), the National Key R&D Program of China ( 2017YFC0908600 ), the Zhejiang Provincial Natural Science Foundation of China ( LY18H300003 ). Scientific Research Fund of Zhejiang Provincial Education Department ( Y201430444 ). Publisher Copyright: {\textcopyright} 2019 Elsevier B.V.",

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

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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

N1 - Funding Information: We acknowledge financial support from the National Natural Science Foundation of China (Grant 81573389 ), the National Key R&D Program of China ( 2017YFC0908600 ), the Zhejiang Provincial Natural Science Foundation of China ( LY18H300003 ). Scientific Research Fund of Zhejiang Provincial Education Department ( Y201430444 ). Publisher Copyright: © 2019 Elsevier B.V.

PY - 2019/10/24

Y1 - 2019/10/24

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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).

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

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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