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Recombinant Proteins> | The BKPyV qPCR and phocine herpesvirus PCR were duplexed for DNA quality and potential PCR inhibition monitoring. Furthermore, the BKPyV qPCR was validated to detect BKPyV genotypes I–IV. qPCR reactions were performed in a total volume of 50 μL, containing 25 μL HotStart Taq mastermix (QIAGEN, Hilden, Germany), 0.5 µmol/L of each primer, 0.35 µmol/L BKPyV probe or 0.4 µmol/L HPyV9 probe, and 3.5 mmol/L MgCl2. Reactions were performed by using a CFX96 real-time detection system (Bio-Rad, Hercules, CA, USA) with the following cycle conditions: 15 min at 95°C followed by 45 cycles of amplification (30 s at 95°C; 30 s at 60°C for HPyV9 qPCR and 55°C for BKPyV qPCR; 30 s at 72°C). For quantification, a standard of pGEX 5×3 HPyV9 VP1 plasmid (Genscript, Piscataway, NJ, USA) and of a quantified BKPyV-positive urine sample were used. | Get A Quote |
Several human polyomaviruses of unknown prevalence and pathogenicity have been identified, including human polyomavirus 9 (HPyV9). To determine rates of HPyV9 infection among immunosuppressed patients, we screened serum samples from 101 kidney transplant patients in the Netherlands for HPyV9 DNA and seroreactivity. A total of 21 patients had positive results for HPyV9 DNA; positivity rates peaked at 3 months after transplantation, but the highest viral loads were measured just after transplantation. During 18 months of follow-up, HPyV9 seroprevalence increased from 33% to 46% among transplant patients; seroprevalence remained stable at ≈30% in a control group of healthy blood donors in whom no HPyV9 DNA was d... More