Molecular approaches for HPV genotyping and HPV-DNA physical status

Abstract

Persistent infection with high-risk human papillomavirus (HPV) genotypes is the leading cause of cervical cancer development. To this end several studies have focused on designing molecular assays for HPV genotyping, which are considered as the gold standard for the early diagnosis of HPV infection. Moreover, the tendency of HPV DNA to be integrated into the host chromosome is a determining event for cervical oncogenesis. Thus, the establishment of molecular techniques was promoted in order to investigate the physical status of the HPV DNA and the locus of viral insertion into the host chromosome. The molecular approaches that have been developed recently facilitate the collection of a wide spectrum of valuable information specific to each individual patient and therefore can significantly contribute to the establishment of a personalised prognosis, diagnosis and treatment of HPV-positive patients. The present review focuses on state of the art molecular assays for HPV detection and genotyping for intra-lesion analyses, it examines molecular approaches for the determination of HPV-DNA physical status and it discusses the criteria for selecting the most appropriate regions of viral DNA to be incorporated in HPV genotyping and in the determination of HPV-DNA physical status.

Cervical cancer risk profiling: molecular biomarkers predicting the outcome of hrHPV infection

“…Although the prevalence of specific hrHPV types varies geographically, several epidemiological studies have shown that the most frequent genotype in invasive cervical cancer is HPV16, followed by HPV18, with both accounting 70% of all cases [10,40,41,54] Figure 2. HPV45 is rarely found in women with normal cytology or low-grade lesions compared to HPV16, but it is the third most common HPV genotype in invasive cervical cancer [55].…”

“…Moreover, the American Society for Colposcopy and Cervical Pathology proposes that positive hrHPV women with normal cytopathological test should undergo HPV genotyping analysis, instead of waiting to be rescreened after one year. This will make it possible to triage women with a low risk of progressing to CIN3/cancer from those with a higher risk [10]. Additionally, Ebisch et al suggested that the combination of HPV16/18 genotyping with cytology testing may improve the referral rate and specificity for detecting CIN3+ lesions, maintaining the sensitivity during triage [57].…”

“…In addition to hrHPV DNA testing, which only gives information about the virus presence, a significant interest is focused on the use of viral mRNA as a biomarker, particularly the upregulation of E6/E7 oncoproteins, to further understand the infection state [10,72].…”

“…In the upper epithelial layer, the E4 positive cells leave the cell cycle and express the two structural proteins, L1 and L2. These particles allow viral genome assembly, packaging, and releasing of new infective virions [6,[8][9][10][11].…”

Association of p16 (CDKN2A) polymorphisms with the development of HPV16‐related precancerous lesions and cervical cancer in the Greek population

“…The Alphapapillomaviruses can be further sub‐divided into high‐risk (HR) genotypes and low‐risk (LR) genotypes considering their epidemiological association with neoplastic disease . Persistent infection with high‐risk HPV genotypes is regarded as the most crucial risk factor for cervical cancer development . Cancer of the uterine cervix is the third most common malignancy in women in terms of incidence and mortality rates and it is the second leading cause of cancer‐associated deaths, worldwide, with higher frequency in low‐income countries .…”