Reliable quantification of rhinovirus species C using real-time PCR

•Developed and validated 4 in-house assays to reliably determine viral load of several different RV-C genotypes in nasal samples.•Complete homology between assay and target sequence is the most accurate method to determine viral load.•Three or more mismatches between the probe and target sequence results in substantially inaccurate viral load.•These assays facilitate accurate investigations into the role of RV-C load in disease outcome.