(A-D) Quantitative Real time PCR of HPV. Quantitative Real time PCR of GP5+/GP6+ to detect presence of HPV infection in blood and tumour biopsy DNA from breast cancer patients. Genomic DNA isolated from blood and tumour tissue biopsies from breast cancer patients were amplified using GP5+/GP6+ L1 consensus PCR primers using Biorad iQ SYBR Green supermix as described in Methods. Ten fold dilution series of WHO HPV16 international standards in the 4 log dynamic range (5 × 104-100) in C33a DNA diluents were used as reference for generation of standard curves. (A) DNA amplification; (B) Standard curve analysis showing efficiency of reaction and calculation of viral copy number; (C) Melt curve analysis showing specificity of HPV16 amplicon and (D) Post real time PCR electrophoresis run of standards and analyzed sample amplicons.