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  Indian J Med Microbiol
 

Figure 2. Determination of physical state HPV16 genome in cervical precancer and cancer tissue biopsies by HPV16 E2 and HPV16 E6 amplification. HPV16 E2 and E6 open reading frame (ORFs) were amplified using specific primers, and E2:E6 densitometric ratio was determined as described in 'Methods'. Vector-free HPV16 full length plasmid harbouring intact E2 and E6 region was used as positive control and as a reference for normalization of E2:E6 ratio in clinical samples. Upper panels showing gel photograph of HPV16 E2 amplification (1139bp) and lower panels show HPV16 E6 amplification (506bp) in DNA of vector control and clinical samples. Normalized E2:E6 ratio of the samples is indicated at the bottom of the respective lane. Normalized E2:E6 ratio = 0 represents completely integrated HPV16 genome, value = 1 represent episomal viral genome, and values between 0 and 1 indicate mixed form of HPV16 genome. M, φX174 HaeIII-digested molecular weight marker; P, HPV16 plasmid; N, PCR negative control (C33a genomic DNA); L1-L3, LSIL cases; H1- H3, HSIL cases; C1- C4, invasive cancer cases

Figure 2. Determination of physical state HPV16 genome in cervical precancer and cancer tissue biopsies by HPV16 E2 and HPV16 E6 amplification. HPV16 E2 and E6 open reading frame (ORFs) were amplified using specific primers, and E2:E6 densitometric ratio was determined as described in 'Methods'. Vector-free HPV16 full length plasmid harbouring intact E2 and E6 region was used as positive control and as a reference for normalization of E2:E6 ratio in clinical samples. Upper panels showing gel photograph of HPV16 E2 amplification (1139bp) and lower panels show HPV16 E6 amplification (506bp) in DNA of vector control and clinical samples. Normalized E2:E6 ratio of the samples is indicated at the bottom of the respective lane. Normalized E2:E6 ratio = 0 represents completely integrated HPV16 genome, value = 1 represent episomal viral genome, and values between 0 and 1 indicate mixed form of HPV16 genome. M, φX174 HaeIII-digested molecular weight marker; P, HPV16 plasmid; N, PCR negative control (C33a genomic DNA); L1-L3, LSIL cases; H1- H3, HSIL cases; C1- C4, invasive cancer cases