title: PrimRglo: a multiplexable quantitative real-time polymerase chain reaction system for nucleic acid detection.
abstract: We report the development of a new real-time polymerase chain reaction (PCR) detection system that uses oligonucleotide "tagged" PCR primers, a fluorophore-labeled "universal" detection oligonucleotides, and a complementary quenching oligonucleotide. The fluorescence signal decreases as PCR product accumulates due to the increase in detection/quencher hybrid formation as the tagged primer is consumed. We use plasmids containing the influenza A matrix gene and the porA and ctrA genes of Neisseria meningitidis as targets for developing the system. Cycle threshold (Ct) values were generated, and the sensitivity of the new system (dubbed "PrimRglo") compared favorably with the commonly used SYBR green and Taqman detection systems and, unlike the latter system, does not require the design of a new dual-labeled detection oligonucleotide for each new target sequence.
authors.affiliation: Biochip Innovations, Mount Gravatt, Queensland 4122, Australia.
authors.forename: Richard
authors.initials: R
authors.lastname: Lai
authors.forename: Fang
authors.initials: F
authors.lastname: Liang
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: Ross T
authors.initials: RT
authors.lastname: Barnard
authors.forename: Simon R
authors.initials: SR
authors.lastname: Corrie
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000138:chemical synthesis; D015342:DNA Probes / Q000138:chemical synthesis; D004268:DNA-Binding Proteins / Q000235:genetics; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009696:Nucleic Acids / Q000032:analysis*; D010957:Plasmids; D018272:Porins / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D013194:Staining and Labeling; D014763:Viral Matrix Proteins / Q000235:genetics
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; C488936:M1 protein, Influenza A virus; D009696:Nucleic Acids; D018272:Porins; D014763:Viral Matrix Proteins; C057699:porin protein, Neisseria