Razvoj širokospektralne verižne reakcije s polimerazo za dokazovanje virusov iz rodu Mupapillomavirus

Vončina, Andraž (author), Poljak, Mario (mentor)

Človeški papilomavirusi (HPV) so zelo majhni in raznoliki virusi s krožnim DNA genomom, ki so povezani z nastankom različnih benignih in malignih sprememb kože in sluznic. V rod Mupapillomavirus (Mu-PV) trenutno uvrščamo tri genotipe HPV (HPV-1, HPV-63 in HPV-204). V magistrskem delu smo razvili in uvedli občutljivo in specifično rodovno-značilno širokospektralno verižno reakcijo s polimerazo (PCR), ki omogoča pomnoževanje 375-bp dolgega dela gena E1 vseh treh uradno priznanih genotipov HPV iz rodu Mu-PV in odkrivanje sorodnih potencialno novih genotipov HPV. Novo-razvito reakcijo PCR smo uporabili za testiranje DNA izolatov, osamljenih iz brisov površine kožnih bradavic (n=29) in brisov površine turške savne (n=55), ter tako pridobili ene izmed prvih podatkov o prevalenci Mu-PV na površini kože in v okolju. Prisotnost HPV-DNA smo dokazali v 58,6 % brisov histološko potrjenih kožnih bradavic ter v 40 % brisov javnih površin. V enem brisu kožne bradavice smo odkrili prisotnost potencialno novega genotipa HPV, ki je na podlagi delnega nukleotidnega zaporedja gena E1 filogenetsko najbolj soroden nedavno odkritemu genotipu HPV-204.

• Language: Slovenian
• Keywords: virusi, človeški papilomavirusi, Mupapillomavirusi, HPV, HPV-1, HPV-63, HPV-204, brisi, kožne bradavice, savne, diagnostične metode, molekularne tehnike, PCR, širokospektralni PCR, sekveniranje
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [A. Vončina]
• Year: 2017
• PID: 20.500.12556/RUL-92895
• UDC: 578.7:616-006.5:577.2.083
• COBISS.SI-ID: 4794744
• Publication date in RUL: 07.07.2017

Secondary language

• Language: English
• Title: Development of the broad-spectrum polymerase chain reaction for detection of human papillomaviruses from the Mupapillomavirus genus

Abstract

Human papillomaviruses (HPVs) are a group of small and diverse viruses with a circular DNA genome, which are associated with the development of various benign and malignant neoplasms of the skin and mucosa. To date only three officially recognized HPV types cluster to the Mupapillomavirus (Mu-PV) genus (HPV-1, HPV-63 in HPV-204). In the present study we have developed and introduced a sensitive and specific broad-spectrum polymerase chain reaction (PCR), which enables amplification of the 375-bp fragment of E1 genes of all three Mu-PV types. We have used this newly developed PCR for testing DNA isolates extracted from surface swabs of skin warts (n=29) and swabs from the surface of Turkish sauna (n=55). Consequently, we have obtained one of the first data on the prevalence of Mu-PVs on the skin and in the environment. The presence of HPV-DNA was determined in 58.6 and 40 % of swabs of histologically confirmed skin warts and public surfaces, respectively. In one swab of the skin wart, we have identified a presence of a potentially novel HPV type, which is based on the partial nucleotide sequence of the E1 gene, phylogenetically most closely related to the newly described HPV-204.

• Keywords: viruses, human papillomaviruses, Mupapillomaviruses, HPV, HPV-1, HPV-63, HPV-204, swabs, skin warts, sauna, diagnostic methods, molecular techniques, PCR, broad-spectrum PCR, sequencing