High-throughput analysis of molecular markers for het HPV-induced carcinogenesis

Datum: 21 januari 2014

Locatie: UAntwerp - Campus Drie Eiken - Promotiezaal - Universiteitsplein 1 - 2610 Wilrijk

Tijdstip: 16 uur

Organisatie / co-organisatie: Faculty of Medicine and Health Sciences

Promovendus: Isabel Micalessi

Promotor: Prof. J.P. Bogers and Dr. G. Boulet

Korte beschrijving: PhD defence Isabel Micalessi - Faculty of Medicine and Health Sciences

Abstract: The pivotal role of HR-HPV in the carcinogenesis has paved the way for HPV-based prevention strategies, i.e. HPV vaccination and screening based on HPV detection. Cervical carcinoma is the most prevalent HPV-related cancer, but screening has significantly reduced its incidence. Overwhelming evidence proves that primary HR-HPV DNA testing is a more sensitive screening method than conventional cytology. However, due to the low specificity of HPV DNA detection, efficient screening requires alternative tests with a better specificity or highly specific triage strategies. New assays based on molecular markers may provide a better diagnostic accuracy and could also be applied in the future screening of other HPV-associated cancers. Besides the application as a screening tool, the molecular analysis of HPV-related markers is valuable for epidemiological studies, vaccine trials and fundamental research purposes.
The implementation of molecular markers in routine laboratory testing not only depends on their diagnostic accuracy, but also on their high-throughput applicability. The research described in this thesis demonstrates that HPV-associated molecular markers, such as HPV DNA, type, load and cellular proteins are amenable to high-throughput analysis in the context of screening or for HPV research purposes. Multiplexing of type-specific primers combined with fluorescent probes for real-time detection allows high-throughput type-specific HPV detection and quantification with excellent turnaround times. High-throughput HPV detection has also been accomplished by multiplexing SPF10 consensus primers in a single-tube real-time PCR assay. This approach considerably reduces the workload and cost associated with HPV typing by the LiPA assay. The high-throughput detection of cellular proteins associated with progressive HPV infections to identify dysplastic cells has proven to be feasible with the BD Pathway high-content cell analyser. However, this methodology should be further clinically evaluated following the five-phase framework for biomarker validation with the purpose of use in screening.