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

Author

Liang F, Arora N, Zhang KL, Yeh DC, Lai R, Pearson D, Barnett G, Whiley D, Sloots T, Corrie SR, and Barnard RT

Subjects
Base Sequence, DNA Primers chemistry, DNA Primers genetics, DNA, Bacterial genetics, DNA, Bacterial isolation & purification, DNA, Viral genetics, DNA, Viral isolation & purification, Escherichia coli, Fluorescent Dyes chemistry, Influenza A virus genetics, Molecular Sequence Data, Neisseria meningitidis genetics, Porins genetics, RNA, Viral genetics, RNA, Viral isolation & purification, Sensitivity and Specificity, Multiplex Polymerase Chain Reaction methods, Real-Time Polymerase Chain Reaction methods
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.

Published
2013
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