A new, multiplex, quantitative real-time polymerase chain reaction system for nucleic acid detection and quantification

Fang Liang, Neetika Arora, Kang Liang Zhang, David Che Cheng Yeh, Richard Lai, Darnley Pearson, Graeme Barnett, David M Whiley, Theo Sloots, Simon Robert Corrie, Ross Thomas Barnard

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

4 Citations (Scopus)

Abstract

Quantitative real-time polymerase chain reaction (qPCR) has emerged as a powerful investigative and diagnostic tool with potential to generate accurate and reproducible results. qPCR can be designed to fulfil the four key aspects required for the detection of nucleic acids: simplicity, speed, sensitivity, and specificity.This chapter reports the development of a novel real-time multiplex quantitative PCR technology, dubbed PrimRglo™, with a potential for high-degree multiplexing. It combines the capacity to simultaneously detect many viruses, bacteria, or nucleic acids, in a single reaction tube, with the ability to quantitate viral or bacterial load. The system utilizes oligonucleotide-tagged PCR primers, along with complementary fluorophore-labelled and quencher-labelled oligonucleotides. The analytic sensitivity of PrimRglo technology was compared with the widely used Taqman® and SYBR green detection systems.
Original languageEnglish
Title of host publicationNucleic Acid Detection
Subtitle of host publicationMethods and Protocols
EditorsDmitry M. Kolpashchikov, Yulia V. Gerasimova
Place of PublicationNew York NY USA
PublisherHumana Press
Pages51 - 68
Number of pages18
Edition1
ISBN (Electronic)9781627035354
ISBN (Print)9781627035347
DOIs
Publication statusPublished - 2 Jul 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1039
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Quantitative PCR
  • Multiplexed PCR
  • Primer design
  • PrimRglo
  • Nucleic acid

Cite this

Liang, F., Arora, N., Zhang, K. L., Yeh, D. C. C., Lai, R., Pearson, D., ... Barnard, R. T. (2013). A new, multiplex, quantitative real-time polymerase chain reaction system for nucleic acid detection and quantification. In D. M. Kolpashchikov, & Y. V. Gerasimova (Eds.), Nucleic Acid Detection : Methods and Protocols (1 ed., pp. 51 - 68). (Methods in Molecular Biology; Vol. 1039). New York NY USA: Humana Press. https://doi.org/10.1007/978-1-62703-535-4_4
Liang, Fang ; Arora, Neetika ; Zhang, Kang Liang ; Yeh, David Che Cheng ; Lai, Richard ; Pearson, Darnley ; Barnett, Graeme ; Whiley, David M ; Sloots, Theo ; Corrie, Simon Robert ; Barnard, Ross Thomas. / A new, multiplex, quantitative real-time polymerase chain reaction system for nucleic acid detection and quantification. Nucleic Acid Detection : Methods and Protocols. editor / Dmitry M. Kolpashchikov ; Yulia V. Gerasimova. 1. ed. New York NY USA : Humana Press, 2013. pp. 51 - 68 (Methods in Molecular Biology).
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Liang, F, Arora, N, Zhang, KL, Yeh, DCC, Lai, R, Pearson, D, Barnett, G, Whiley, DM, Sloots, T, Corrie, SR & Barnard, RT 2013, A new, multiplex, quantitative real-time polymerase chain reaction system for nucleic acid detection and quantification. in DM Kolpashchikov & YV Gerasimova (eds), Nucleic Acid Detection : Methods and Protocols. 1 edn, Methods in Molecular Biology, vol. 1039, Humana Press, New York NY USA, pp. 51 - 68. https://doi.org/10.1007/978-1-62703-535-4_4

A new, multiplex, quantitative real-time polymerase chain reaction system for nucleic acid detection and quantification. / Liang, Fang; Arora, Neetika; Zhang, Kang Liang; Yeh, David Che Cheng; Lai, Richard; Pearson, Darnley; Barnett, Graeme; Whiley, David M; Sloots, Theo; Corrie, Simon Robert; Barnard, Ross Thomas.

Nucleic Acid Detection : Methods and Protocols. ed. / Dmitry M. Kolpashchikov; Yulia V. Gerasimova. 1. ed. New York NY USA : Humana Press, 2013. p. 51 - 68 (Methods in Molecular Biology; Vol. 1039).

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

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AU - Liang, Fang

AU - Arora, Neetika

AU - Zhang, Kang Liang

AU - Yeh, David Che Cheng

AU - Lai, Richard

AU - Pearson, Darnley

AU - Barnett, Graeme

AU - Whiley, David M

AU - Sloots, Theo

AU - Corrie, Simon Robert

AU - Barnard, Ross Thomas

PY - 2013/7/2

Y1 - 2013/7/2

N2 - Quantitative real-time polymerase chain reaction (qPCR) has emerged as a powerful investigative and diagnostic tool with potential to generate accurate and reproducible results. qPCR can be designed to fulfil the four key aspects required for the detection of nucleic acids: simplicity, speed, sensitivity, and specificity.This chapter reports the development of a novel real-time multiplex quantitative PCR technology, dubbed PrimRglo™, with a potential for high-degree multiplexing. It combines the capacity to simultaneously detect many viruses, bacteria, or nucleic acids, in a single reaction tube, with the ability to quantitate viral or bacterial load. The system utilizes oligonucleotide-tagged PCR primers, along with complementary fluorophore-labelled and quencher-labelled oligonucleotides. The analytic sensitivity of PrimRglo technology was compared with the widely used Taqman® and SYBR green detection systems.

AB - Quantitative real-time polymerase chain reaction (qPCR) has emerged as a powerful investigative and diagnostic tool with potential to generate accurate and reproducible results. qPCR can be designed to fulfil the four key aspects required for the detection of nucleic acids: simplicity, speed, sensitivity, and specificity.This chapter reports the development of a novel real-time multiplex quantitative PCR technology, dubbed PrimRglo™, with a potential for high-degree multiplexing. It combines the capacity to simultaneously detect many viruses, bacteria, or nucleic acids, in a single reaction tube, with the ability to quantitate viral or bacterial load. The system utilizes oligonucleotide-tagged PCR primers, along with complementary fluorophore-labelled and quencher-labelled oligonucleotides. The analytic sensitivity of PrimRglo technology was compared with the widely used Taqman® and SYBR green detection systems.

KW - Quantitative PCR

KW - Multiplexed PCR

KW - Primer design

KW - PrimRglo

KW - Nucleic acid

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U2 - 10.1007/978-1-62703-535-4_4

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M3 - Chapter (Book)

SN - 9781627035347

T3 - Methods in Molecular Biology

SP - 51

EP - 68

BT - Nucleic Acid Detection

A2 - Kolpashchikov, Dmitry M.

A2 - Gerasimova, Yulia V.

PB - Humana Press

CY - New York NY USA

ER -

Liang F, Arora N, Zhang KL, Yeh DCC, Lai R, Pearson D et al. A new, multiplex, quantitative real-time polymerase chain reaction system for nucleic acid detection and quantification. In Kolpashchikov DM, Gerasimova YV, editors, Nucleic Acid Detection : Methods and Protocols. 1 ed. New York NY USA: Humana Press. 2013. p. 51 - 68. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-535-4_4