A home-made method for human papillomavirus detection in head and neck cancers

Fazzi, M. (Maran) (2017) A home-made method for human papillomavirus detection in head and neck cancers. thesis, Medicine.

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Abstract

Abstract: Since the beginning of this century it has become clear that, besides cervical cancer, human papillomavirus (HPV) is causing a subset of other cancers, among which are oropharyngeal squamous cell carcinomas (OPSCC). The incidence of HPV related OPSCCs is rising and it is estimated that its incidence will exceed the one of HPV related cervical cancer in 2020. HPV related OPSCCs respond better to treatment and have a better prognosis than HPV negative OPSCCs. That is why it is important to be able to reliably establish tumor HPV status, in order to, in the future, apply de-intensification of treatment. There is need for a test that is specially designed for head and neck cancers, as all current tests are developed for the cervix and lack specificity. There is a gold standard for fresh tumor samples, but there is need for a suitable test on formalin-fixed, paraffin-embedded (FFPE) tumor samples, which we aimed to find. We tested a home-made method, consisting of a home-made primer mix for HPV detection by testing for E7 oncogene DNA of HPV type 16 and 18 by using quantative real-time polymerase chain reaction (qPCR). We tested this method on 72 fresh head and neck squamous cell carcinoma (HNSCC) samples and 30 OPSCC FFPE samples and compared it to the gold standard and the commercially available Digene Hybrid Capture 2 assay and Anyplex II HPV HR Detection methods, which were tested respectively on 108, 108 and 85 fresh samples. Performance of the different methods was calculated using kappa’s coefficient for inter-rater agreement between each test and the gold-standard. The home-made method on fresh samples scored in the same category as the Digene method with a κ value of 0.83 compared to 0.91 and performed better than the Anyplex method, which scored 0.75. The home-made method performed rather poorly on FFPE samples with κ values of 0.21 and 0.18, depending on replication cycle cut-off values for qPCR. It can be concluded that the home-made method works on fresh samples, but does not yet perform sufficiently on FFPE samples.

Item Type:	Thesis (Thesis)
Supervisor name:	Faculty supervisor: and Dikkers, F.G. MD-PhD
Supervisor name:	Second supervisor: and Bussu, Francesco MD–PhD
Faculty:	Medical Sciences
Date Deposited:	25 Jun 2020 11:04
Last Modified:	25 Jun 2020 11:04
URI:	https://umcg.studenttheses.ub.rug.nl/id/eprint/2405

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