title: A new, multiplex, quantitative real-time polymerase chain reaction system for nucleic acid detection and quantification.
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.
authors.affiliation: Australian Infectious Diseases Research Centre, School of Chemistry and Molecular Biosciences, University of Queensland, St. Lucia, QLD, Australia.
authors.forename: Fang
authors.initials: F
authors.lastname: Liang
authors.forename: Neetika
authors.initials: N
authors.lastname: Arora
authors.forename: Kang Liang
authors.initials: KL
authors.lastname: Zhang
authors.forename: David Che Cheng
authors.initials: DC
authors.lastname: Yeh
authors.forename: Richard
authors.initials: R
authors.lastname: Lai
authors.forename: Darnley
authors.initials: D
authors.lastname: Pearson
authors.forename: Graeme
authors.initials: G
authors.lastname: Barnett
authors.forename: David
authors.initials: D
authors.lastname: Whiley
authors.forename: Theo
authors.initials: T
authors.lastname: Sloots
authors.forename: Simon R
authors.initials: SR
authors.lastname: Corrie
authors.forename: Ross T
authors.initials: RT
authors.lastname: Barnard
mesh_terms: D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli; D005456:Fluorescent Dyes / Q000737:chemistry; D009980:Influenza A virus / Q000235:genetics; D008969:Molecular Sequence Data; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009345:Neisseria meningitidis / Q000235:genetics; D018272:Porins / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D004279:DNA, Viral; D005456:Fluorescent Dyes; D018272:Porins; D012367:RNA, Viral; C057699:porin protein, Neisseria