title: Polymerase chain displacement reaction.
abstract: Quantitative PCR assays are now the standard method for viral diagnostics. These assays must be specific, as well as sensitive, to detect the potentially low starting copy number of viral genomic material. We describe a new technique, polymerase chain displacement reaction (PCDR), which uses multiple nested primers in a rapid, capped, one-tube reaction that increases the sensitivity of normal quantitative PCR (qPCR) assays. Sensitivity was increased by approximately 10-fold in a proof-of-principle test on dengue virus sequence. In PCDR, when extension occurs from the outer primer, it displaces the extension strand produced from the inner primer by utilizing a polymerase that has strand displacement activity. This allows a greater than 2-fold increase of amplification product for each amplification cycle and therefore increased sensitivity and speed over conventional PCR. Increased sensitivity in PCDR would be useful in nucleic acid detection for viral diagnostics.
authors: Oxitec Ltd, Abingdon, Oxfordshire, UK.
authors: Claire L
authors: CL
authors: Harris
authors: Irma J
authors: IJ
authors: Sanchez-Vargas
authors: Ken E
authors: KE
authors: Olson
authors: Luke
authors: L
authors: Alphey
authors: Guoliang
authors: G
authors: Fu
mesh_terms: D000330:Aedes / Q000821:virology; D000818:Animals; D017931:DNA Primers; D018076:DNA, Complementary / Q000032:analysis / Q000378:metabolism; D003716:Dengue Virus / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000302:isolation & purification / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D012367:RNA, Viral