Genotipizacija humanih papilomavirusa

master's thesis

• Title (english): Human papillomavirus genotyping
• Author: Ana Škara
• Mentor: Snježana Židovec-Lepej (mentor)
• Mentor: Nada Oršolić (sumentor)
• Committee member: Snježana Židovec-Lepej (predsjednik povjerenstva)
• Committee member: Nada Oršolić (član povjerenstva)
• Committee member: Željka Vidaković Cifrek (član povjerenstva)
• Committee member: Mirjana Kalafatić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2009, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Humani papilomavirusi (rod: Alpha-papillomavirus, porodica: Papillomaviridae) su heterogena skupina DNK virusa koji imaju značajnu ulogu u nastanku benignih i malignih novotvorina višeslojnog pločastog epitela. Cilj ovog rada bio je analizirati učestalost detekcije genotipova HPV-a koji su povezani s visokim odnosno niskim rizikom od nastanka raka vrata maternice u obriscima cerviksa žena koje se kontroliraju u ambulanti za urogenitalne infekcije kliničke bolnice. U dijelu uzoraka koji su bili pozitivni u kvalitativnom testu na HPV DNK odredila sam pojedinačne genotipove HPV-a i analizirala njihovu distribuciju. Metodom reverzne hibridizacije (hybrid capture 2 test) određivala sam prisutnost genotipova HPV-a koji su povezani s visokim odnosno niskim rizikom od nastanka raka vrata maternice. U radu sam koristila grupne hibridizacijske probe za visokorizične genotipove HPV-a (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68) i niskorizične genotipove (6, 11, 42, 43, 44). Identifikaciju pojedinih genotipova HPV-a u uzorcima koji su prethodno bili pozitivni u kvalitativnom hibridizacijskom testu na HPV DNK odredila sam kombinacijom PCR-a i reverzne hibridizacije. Nakon izolacije DNK, PCR reakcijom umnožen je dio L1 regije HPV DNK a nakon toga je provedena reverzna hibridizacija primjenom oligonukleotidnih proba vezanih na nitrocelulozne membrane. Rezultati kvalitativne analize visokorizičnih i niskorizičnih genotipova HPV-a pokazali su da je HPV DNK bila detektirana u 178 od 429 ispitanica (41.4%). Individualna genotipizacija HPV-a pokazala je da su HPV-16 i HPV-31 najučestaliji genotipovi HPV-a u uzorcima koje sam analizirala u ovom radu.

Abstract (english)

Human papillomaviruses are a heterogenous group of DNA viruses that have significant role in appearance of benign and malignant neoplasm in multilayer epithelium. The purpose of this work was to analyze a frequency in detection of HPV genotypes related to high-risk and lowrisks of cervix cancer formation in cervical swabs. The specimens were collected from women attending the Outpatient Clinics for Urogenital infections of University Hospital for Infectious Diseases "Dr Fran Mihaljevic". For specimens that were positive on the HPV DNA qualitative test, individual HPV genotypes were determined and their distribution was analysed. Using a Hybrid Capture 2 test, the presence of HPV genotypes associated with high-risk or low-risk for the development of cervical cancer was analysed. Hybridization probes specific for high-risk HPV genotypes (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68), and low-risk genotypes (HPV 6, 11, 42, 43, 44) were used in this study. In specimens that were previously positive by HPV DNA qualitative hybridization test, individual HPV genotypes were identified by combining PCR and reverse hybridization. Following DNA isolation, a part of L1 region of the HPV DNA was amplified by PCR reaction. In the next step, the reverse hybridization and oligonucleotide probes were used. These probes were attached to nitrocellulose membranes. The results of the qualitative analysis of high-risk and low-risk HPV genotypes showed that HPV DNA was detected in 178 of 429 women (41.4%). Individual HPV genotyping showed that HPV-16 and HPV-31 were the most frequent HPV genotypes in analyzed specimens.

• Keywords: humani papilomavirusi; rak vrata maternice; cervikalna intraepitelijalna neoplazija; reverzna hibridizacija; lančana reakcija polimeraze
• Keywords (english): human papillomaviruses; cervical cancer; cervical intraepithelial neoplasia; reverse hybridization; polymerase chain reaction
• Language: croatian
• URN:NBN: urn:nbn:hr:217:180586
• Study programme: Title: Biology; specializations in: Molecular biology; Course: Molecular biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra biologije (magistar/magistra biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2018-03-21 10:05:02