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Molekularna opredelitev povzročiteljev navadnih kožnih bradavic
ID Skubic, Lucijan (Author), ID Poljak, Mario (Mentor) More about this mentor... This link opens in a new window

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Abstract
Človeški papilomavirusi (HPV) so etiološko povezani z nastankom številnih benignih kožnih novotvorb, med katerimi so najpogostejše navadne kožne bradavice (verrucae vulgares). Namen naše raziskave je bil, da z različnimi molekularnimi metodami prvič doslej zanesljivo določimo HPV povzročitelja v vsaj 95 % primerov navadnih kožnih bradavic. V sklopu vzpostavljenega diagnostičnega protokola smo z uporabo široko-spektralne verižne reakcije s polimerazo (PCR), sekveniranja po Sangerju in dveh predhodno opisanih ter sedmih novo razvitih tipsko-značilnih kvantitativnih PCR v realnem času (qPCR) v kombinaciji s qPCR za pomnoževanje dela gena za človeški beta-globin testirali 132 histološko-potrjenih navadnih kožnih bradavic in določili etiološko vlogo vseh zaznanih genotipov HPV na podlagi ocene virusnega bremena. Skupno smo kot povzročitelje opredelili 12 različnih genotipov HPV v 126/130 (96,9 %) HPV pozitivnih navadnih kožnih bradavic, med katerimi so prevladovali HPV27 (27,7 %), HPV57 (26,1 %), HPV4 (14,6 %), HPV2 (13,1 %) in HPV65 (7,7 %). Z metagenomskim sekveniranjem naslednje generacije v bradavicah brez uspešne etiološke opredelitve nismo zaznali HPV DNA. S tipsko-značilnimi protokoli digitalnega PCR smo z višjo stopnjo natančnosti kvantifikacije tarčnih genotipov HPV v testiranih bradavicah potrdili enake povzročitelje kot s tipsko-značilnimi qPCR, brez statistično značilnih razlik v vrednostih virusnih bremen med obema metodama, pri čemer je imel vzpostavljen diagnostični protokol širše dinamično območje.

Language:Slovenian
Keywords:človeški papilomavirusi, navadne kožne bradavice, povzročitelji, molekularna opredelitev, virusno breme
Work type:Doctoral dissertation
Typology:2.08 - Doctoral Dissertation
Organization:BF - Biotechnical Faculty
Publisher:[L. Skubic]
Year:2023
PID:20.500.12556/RUL-148218 This link opens in a new window
UDC:578.72:616-006.5:577.2.083
COBISS.SI-ID:160473859 This link opens in a new window
Publication date in RUL:03.08.2023
Views:1535
Downloads:66
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Secondary language

Language:English
Title:Molecular characterization of etiological agents of common skin warts
Abstract:
Human papillomaviruses (HPVs) are etiologically associated with various benign skin neoplasms, of which common warts (verrucae vulgares) are the most prevalent. The aim of our study was to reliably determine for the first time the etiological agent in at least 95% of common warts by a combination of different molecular methods. Within the established diagnostic protocol, we used broad-spectrum polymerase chain reactions (PCRs) followed by Sanger sequencing, two previously described and seven newly developed type-specific quantitative real-time PCRs (qPCRs) coupled with the human beta-globin gene qPCR to analyse 132 histologically confirmed common warts and to determine the etiologic role of all detected HPVs based on viral load estimation. A total of 12 different causative HPV types were determined in 126/130 (96.9%) HPV-positive common warts, with HPV27 (27.7%), HPV57 (26.1%), HPV4 (14.6%), HPV2 (13.1%), and HPV65 (7.7%) being the most prevalent. Metagenomic next-generation sequencing showed that HPV DNA was not present in any of the warts without a determined etiological agent. Using the type-specific digital PCRs, we confirmed the same etiologic agents previously identified by type-specific qPCRs in the warts tested, with higher precision in quantifying target HPV types and no statistically significant differences in viral loads, whereas the established diagnostic protocol had a wider dynamic range.

Keywords:human papillomaviruses, common skin warts, etiological agents, molecular characterization, viral load

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