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Updated: Aug 15, 2026
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Novel Sequence Discovery by Subtractive Genomics
Published on: January 25, 2019
Sequence variation in primer targets affects the accuracy of viral quantitative PCR
David M Whiley1, Theo P Sloots
1Clinical Virology Research Unit, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Qld 4029, Australia.
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.
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