Sequence variation in primer targets affects the accuracy of viral quantitative PCR.
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.
Whiley
David M
DM
Clinical Virology Research Unit, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Qld 4029, Australia.
Sloots
Theo P
TP
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*
D017931:DNA Primers