3.1.2. Sensitivity and specificity of the new MCPyV quantitative
assay
The qPCR assay developed for the quantification of MCPyV was
shown to be specific both by “in silico” sequence analysis of the
primers and the probe considering nucleotide sequence databases
(NCBI BLAST) and by experimental assays. No false positive results
were detected due to cross-reactivity with non-target DNA from
the viruses contained in the different plasmid constructs assayed
(human polyomaviruses JCPyV, BKPyV, KIPyV and WUPyV and
animal polyomaviruses OPyV and BPyV). A total of 20 and 10 DNA
genome copies were detected in 100% and 80% of the performed
qPCR reactions, respectively. Sensitivity did not vary even when
high levels of exogenous but related viral DNA (samples with high
levels of JCPyV) were added to the test tubes.