Artus cmv rg pcr kit

DNA was extracted from urine, breast milk and vaginal and saliva swabs using an EZ1 DSP virus kit (Qiagen, Hilden, Germany), and from blood using a QIAmp DNA mini kit (Qiagen). HCMV DNA was quantified using an Artus CMV RG PCR kit (Qiagen). Four of the seven gB genotypes (gB1-4) and the two gH genotypes (gH1-2) were genotyped by PCR [15 (link)]. Genotyping of gB (gB1, gB2, gB3, and gB4) and gH (gH1 and gH2) was performed using two multiplex real-time PCR assays. Primers and probes specific for each genotype were designed at the N terminus of gB and gH except for gB3, for which the cleavage region was amplified. However, in this study gB2 and gB3 were considered collectively as gB2/3. Genotypes gB5-7 are not detectable by this PCR assay. The primer and probe sequences were as described previously [15 (link)]. Each multiplex assay contained 10 µL DNA extract, 25 µL HotStar Master mix (Qiagen, Hilden, Germany), 0.3 µM primers, 0.2 µM probes, and 4.5 mM MgCl₂. Template denaturation and activation of HotStarTaq DNA polymerase were achieved by heating for 15 min at 95°C. This was followed by 45 cycles of denaturation at 95°C for 30 s, annealing at 55°C for 30 s, and extension at 72 °C for 30 s. The assay was carried out in a Rotor-Gene Q 5PLEX instrument (Qiagen, Hilden, Germany).