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

6 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

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