title: Sequence variation in primer targets affects the accuracy of viral quantitative PCR.
abstract: This study was designed to assess the effect of sequence mismatches between primers and their targets on viral quantitative PCR. Numerous primers were constructed incorporating various mismatches with a target sequence on the BKV T antigen gene. When using these primers in standard Taqman two-step cycling conditions, as few as two mismatches in a single primer increased cycle threshold values and significantly influenced the calculation of viral load.
authors.affiliation: Clinical Virology Research Unit, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Qld 4029, Australia.
authors.forename: David M
authors.initials: DM
authors.lastname: Whiley
authors.forename: Theo P
authors.initials: TP
authors.lastname: Sloots
mesh_terms: D001739:BK Virus / Q000302:isolation & purification; D020137:Base Pair Mismatch; D017931:DNA Primers; D016133:Polymerase Chain Reaction / Q000379:methods*; D013698:Templates, Genetic; D019562:Viral Load / Q000379:methods*
chemical_list: D017931:DNA Primers