High-risk HPV Detection and Genotyping

High-risk (HR)-HPV DNA in-situ hybridization (ISH) was done using proprietary reagents (Inform HPV III Family 16 Probe [B], Ventana Medical Systems, Inc., USA), which can detect high risk HPV genotypes (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 66) [13 (link)]. HR-HPV(+) cervical specimens were used as the positive control group and HR-HPV(−) cervical specimens were used as the negative control group. The HR-HPV ISH test was scored as positive if there was any blue reaction product that co-localized with cell nuclei [13 (link)]. The staining was reported as positive if it showed diffuse nuclear and cytoplasmic staining or punctate nuclear staining. Staining that was pale and limited to the nucleoli of cells were reported as negative.
Flow-through hybridization was performed for HPV genotyping, using an HBGA-21 GenoArray Diagnostic Kit (HybriBio Biotechnology Ltd. Corp., Chaozhou, Guangdong Province, China) in accordance to the manufacturer’s instructions. The HPV GenoArray helps the identification of 21 HPV genotypes (HPV 6, 11, 16, 18, 31, 33, 35, 39, 42, 43, 44, 45, 51, 52, 53, 56, 58, 59, 66 and 68) and CP8304. Of the 21 genotypes, 5 were classified as low risk (HPV 6, 11, 42, 43, 44) and the remaining HPV genotypes are classified as high-risk [14 (link)].