Figure 3.

Real-time PCR assay performed on tissues (invasive cervical cancer, CIN2+ lesion adjacent to invasive neoplasia, metastatic lymph nodes, non-metastatic lymph nodes, and thyroid), and on liquid-based urinary samples.

Notes: Anyplex II HPV28 detection assay simultaneously detects 28 HPV genotypes. β-globin gene is used as a positive endogen control to identify processed specimens containing substances that may interfere with PCR amplification. Thyroid tissue was used as an external HPV-negative control. (A) Invasive cervical cancer showing positive result for HPV-53. (B) CIN2+ lesion, in which positive results for HPVs 16, 35, 39, 40, 53, 54, 59, 61, 68 and 82, have been found. (C) Metastatic lymph nodes showing HPV-53 positive result. (D) Non-metastatic lymph nodes testing as HPV-negative. (E) Thyroid tissue testing as HPV-negative. (F) Urinary samples in which HPV-53 has been found. (G) Melting curve of HPV-53 control included in the PC3 mix of set B demonstrating the melting curve shape and Tm. Melting profile of samples, in which HPV-53 has been found, showed a Tm of 76.5°C.

Abbreviations: CIN2+, cervical intraepithelial neoplasia grade 2-or-worse; –d(RFU/dT), change in rate of fluorescence units against temperature; IC, internal control; PC3 mix of set B, positive control of set B consisting of a mix of pathogens including those of HPVs 73, 53, 70, 6, and 11; Tm, melting temperature; HPV, human papillomavirus.