Human Papillomavirus (HPV) Genotyping Using Paired Exfoliated Cervicovaginal Cells and Paraffin-Embedded Tissues To Highlight Difficulties in Attributing HPV Types to Specific Lesions

Abstract

Defining type-specific human papillomavirus (HPV) infections within cervical tissues is important for understanding the pathogenesis of cervical neoplasia and assessing the effectiveness of prophylactic vaccines with limited type-specific spectra. We compared HPV DNA-testing results from 146 matched exfoliated-cell and formalin-fixed-tissue specimens collected by cervicovaginal lavage (CVL) within 90 days of each other from women with histologically confirmed cervical intraepithelial lesions (CIN). The CVL specimens were HPV typed using a MY09/11 L1 consensus primer PCR method followed by dot blot hybridization. The tissue specimens were HPV typed using an SPF 10 line probe assay HPV detection system. Of the 146 specimen pairs with evidence of CIN in the tissue, 91.8% were positive for one or more HPV types in both the tissue and cellular specimens. Tissue sections were more likely to be HPV negative (P < 0.01). Typing directly from tissue sections resolved multiple infections detected in exfoliated cells to a single HPV type in only 46.9% of cases. Combined use of both specimen types to attribute lesions to HPV type 16 (HPV-16) and/or -18 led to 43.1% attributed to HPV-16 and/or -18 by both specimen types and 19.9% attributed to HPV-16 and/or -18 by one, but not both, specimen types. Unambiguous attribution of cervical lesions to a single, specific HPV type remains a difficult proposition. Use of multiple specimen types or the development of highly sensitive and robust in situ hybridization HPV-testing methods to evaluate the certainty of attribution of lesions to HPV types might provide insights in future efforts, including HPV vaccine trials.Infection with one of approximately 15 oncogenic types of human papillomavirus (HPV) has been established as a necessary but insufficient cause of cervical cancer (12). Although HPV typing performed by testing DNA extracted from exfoliated cervicovaginal cells is an accurate technique for demonstrating HPV infections, this approach does not meet growing needs to topographically define the specific areas of the cervix that harbor the virus. Among HPV-positive women, 20 to 40% are infected with multiple HPV types. Some HPV types detected by testing exfoliative specimens may reflect vaginal rather than cervical infections, thus underscoring the need for methods that permit localization of viral infection within specific compartments of the cervical mucosa.Clinical trials conducted to test the efficacy of prophylactic vaccines that target two oncogenic HPV types, HPV-16 and HPV-18 (one of the vaccines also targets two nononcogenic HPV types, HPV-6 and HPV-11), have created an immediate need for HPV typing of tissues to colocalize HPV infections and their associated lesions. In most vaccine trials, prevention of cancer precursors (i.e., cervical intraepithelial neoplasia [CIN] grade 2/3) related to HPV-16 and/or -18 is an important end point. Given that prophylactic vaccines are expected to provide minimal protection against untargeted types, it is desirable in trial to accura...

Frequency and Type-distribution of Human Papillomavirus from Paraffin-embedded Blocks of High Grade Cervical Intraepithelial Neoplasia Lesions in Thailand

“…In this study, HPV infection was evaluated by SPF-10-LiPA (INNo-LiPA). The short fragment (65bp) of SPF-10 primers is appropriate amplified the formalin-fixed paraffin-embedded tissue (FFPE) which have a minimal volume and difficult to analyze (Gravitt et al, 2007;van Hamont et al, 2006;Bello et al, 2009).…”

Prevaccination Distribution of Human Papillomavirus Types in Women Attending at Cervical Cancer Screening in Belgium

“…5 Another unresolved issue is the attribution of a lesion to a specific HPV type in case of multiple infections. Persistence of a type over time, sensitive type-specific in situ hybridization, or use of mRNA based tests could clarify this question (51,52). Storage of all residual cellular material, extraction and freezing of DNA, and several feasibility studies involving testing of archived material further enhance the possibility to disentangle the natural history of HPV infections at the type-specific level (53)(54)(55).…”

Human papillomavirus type distribution in 30,848 invasive cervical cancers worldwide: Variation by geographical region, histological type and year of publication

“…However, demonstrating which HPV type is functionally relevant to a tumour, by using techniques such as microdissection and/or detection of E6 or E7 transcripts, remains difficult. 16 It is therefore increasingly challenging to distinguish between clinically relevant and irrelevant HPV infections and thereby to estimate the benefits of tests and vaccines including more or less broad ranges of HPV types. 17 …”