Figure 5

Nested PCR and real-time PCR detecting HPyV6-encoded genes. (A) Sensitivity and specificity of the nested PCR detecting three HPyV6 gene fragments were confirmed. Sequence-validated PCR products of HPyV6 were amplified by the nested PCR. In addition, JCV and BKV-positive controls were examined. (B) Three HPyV6 gene fragments were detected in tissue and serum samples positive for HPyV6 by real-time PCR. A case of MCC (lane 1), Kaposi’s sarcoma (lane 2), and two sera from HIV-1-negative patients (lane 3 and 4) were examined. Average copy numbers of three experiments for HPyV6 ST and human β-actin genes per 100 ng of DNA (tissue samples) or DNA extracted from 1 μL of serum were shown in the bottom. Specific PCR fragments, including large T (LT), VP1, and VP2/3, were not amplified constantly in lane 1-4. Similar results were obtained in a repeated experiment. P: positive control of HPyV6, a DNA sample extracted from a skin biopsy which was confirmed to be HPyV6-positive, N: no DNA.