20.500.12556/RUL-182902 Določanje odpornosti pri virusu influence na zaviralce nevraminidaz Determination of influenza virus resistance to neuraminidase inhibitors Virus influence (FLU) predstavlja enega izmed pomembnejših povzročiteljev okužb dihal in vsako leto predstavljajo znatno breme za javno zdravje. Prenaša se kapljično in primarno prizadene zgornja dihala. Pri ljudeh sta epidemiološko najpomembnejša virus influence A (FluA) in virus influence B (FluB). Želeli smo opredeliti pojavnost različnih vrst in antigenskih različic FLU ter prisotnost genotipske odpornosti na zaviralce nevraminidaz. Zanimalo nas je, ali je pojav odpornosti povezan s starostjo, imunskim statusom, posameznimi antigenskimi različicami virusa ter protivirusnem zdravljenju. V raziskavo smo vključili arhivske vzorce dihal bolnikov s potrjeno prisotnostjo FluA, poslane na Inštitut za mikrobiologijo in imunologijo Medicinske fakultete v Ljubljani. Prisotnost mutacije H275Y smo določali s pomnoževanjem 183 bp dolgega odseka gena za nevraminidazo in analizo alelne diskriminacije z RT-PCR v realnem času. Pri podtipu A(H1N1) smo pomnožili 621 bp in 358 bp dolga fragmenta gena za nevraminidazo, pri podtipu A(H3N2) pa 743 bp dolg odsek, katerim smo nukleotidno zaporedjo določili na genskem analizatorju. Med leti 2022 in 2025 smo potrdili izmenično sezonsko kroženje podtipov A(H1N1) in A(H3N2), medtem ko se je podtip FluB pojavljal le sporadično. Mutacijo H275Y smo pri podtipu A(H1N1) zaznali v manjšem deležu vzorcev, medtem ko pri podtipu A(H3N2) klinično pomembnih mutacij, povezanih z odpornostjo na zaviralce nevraminidaz, nismo zaznali. Mutacija H275Y se je pogosteje pojavljala pri starejših in z oseltamivirjem zdravljenih bolnikih, vendar jasne povezave z imunskim statusom ali zdravljenjem nismo dokazali. Opredelili smo nizko pojavnost genotipske odpornosti ter omejeno kroženje odpornih različic virusa v populaciji. Influenza virus (FLU) is a major cause of respiratory tract infections and remains a substantial public health burden. It is transmitted via respiratory droplets and primarily affects the upper respiratory tract. In humans, influenza virus A (FluA) and influenza virus B (FluB) are the most epidemiologically relevant types. The prevalence of different FLU types and antigenic variants, as well as the presence of genotypic resistance to neuraminidase inhibitors, were investigated. Possible associations between antiviral resistance and age, immune status, specific antigenic virus variants, and antiviral treatment were evaluated. Archived respiratory samples from patients with laboratory-confirmed FluA infection, submitted to the Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, were included in the study. The H275Y mutation was detected by amplification of 183 bp fragment of the neuraminidase gene, followed by allele discrimination analysis using real-time RT-PCR. For subtype A(H1N1), 621 bp and 358 bp fragments of the neuraminidase gene were amplified, while a 743 bp fragment was amplified for subtype A(H3N2). Nucleotide sequences were determined using a genetic analyzer. Between 2022 and 2025, alternating seasonal circulation of subtypes A(H1N1) and A(H3N2) was confirmed, whereas FluB occurred sporadically. The H275Y mutation was detected in a small proportion of A(H1N1) samples, while no clinically relevant resistance-associated mutations were identified in A(H3N2). The H275Y mutation was more frequently observed in older patients and in those treated with oseltamivir; however, no statistically significant association with immune status or treatment was demonstrated. Overall, a low prevalence of genotypic resistance with limited circulation of resistant viral variants in the population was identified. virusi virus influence tipa A zaviralci nevraminidaze genotipska odpornost RT-PCR v realnem času določanje nukleotidnega zaporedja viruses influenza A virus neuraminidase inhibitors genotypic resistance real-time RT-PCR sequencing true false false [K. Rode] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-05-28 07:16:04 2026-05-28 07:16:15 2026-05-29 08:19:26 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 578.2:616.921.5 279829 279768579 57620.pdf 57620.pdf 1 9C13B81A73ECF29BED4EAFBF055D98BB d85d29f6040c9c0034a776adb052df858f7626f636dae83df5467de075f6d631 01c03af8-5a54-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=234553 Biotehniška fakulteta 0 0 0