cCMV Real-time qPCR Primers, Probes, & Controls
The NeoMDx™ cCMV real-time PCR assay uses target sequence-specific primers and TaqMan™ probes (Thermo Fisher Scientific) to amplify and detect cCMV DNA and RPP30, a human gene used as an internal control. The qPCR sample input is extracted DNA from a single dried blood spot (DBS) punch using NeoMDx™ Elution Solution.
The oligonucleotide primers and probes for detection of cCMV are selected from the UL122 gene region (regulatory protein IE2).
The assay also uses a dUTP/UNG carryover prevention system to avoid contamination by PCR products and erroneous results.
There are three DBS kit controls: C1 (CMV negative control), C2 (CMV low positive control), and C3 (CMV high positive control). All three controls are punched and extracted along with the sample before running on the real-time qPCR instrument to ensure the entire workflow is working as intended.