Rare Variants in RPPH1 qPCR Control Assay Binding Sites Result in Incorrect Copy Number Calls

Real-time quantitative PCR (qPCR) using RPPH1 as a reference gene is a standard method for assessment and validation of genomic copy number variations (CNVs). However, variants in the reference amplicon may cause errors, which was investigated here. While conducting CNV validations for birth defects research studies, 13/1,634 specimens with multiple loci that appeared to be present as 3 copies were unexpectedly detected. This apparent trisomy was hypothesized to be an amplification artifact caused by a variant in the RPPH1 amplicon.
Source: Journal of Molecular Diagnostics - Category: Pathology Authors: Tags: Regular Article Source Type: research