20.500.12556/RUL-181421 Klinična opredelitev kemiluminescenčnega imunskega testa za ugotavljanje protiteles proti flavivirusom Clinical evaluation of chemiluminescent immunoassay for the detection of antibodies against flaviviruses Arbovirusne okužbe pri ljudeh lahko povzročijo širok spekter simptomov, od asimptomatskih okužb do življenjsko nevarnih stanj. Zaradi kratke viremije pri arbovirozah pogosto zamudimo diagnostično okno za neposreden dokaz virusa, vendar pa lahko okužbo potrdimo z dokazom specifičnih protiteles z različnimi serološkimi metodami. Glavna omejitev seroloških metod je visoka navzkrižna reaktivnost med flavivirusi, zaradi česar je pogosto težko natančno določiti specifičen patogen. To je spodbudilo razvoj novih seroloških orodij, kot so kemiluminescenčni testi. V magistrsko nalogo smo vključili 108 vzorcev bolnikov s klopnim meningoencefalitisom, dengo in mrzlico Zahodnega Nila ter 12 vzorcev zdravih oseb. Za dokaz posameznih flavivirusnih okužb smo uporabili referenčne laboratorijske teste: Anti-TBE Virus ELISA IgM & IgG, Anti-Dengue Virus ELISA IgM & IgG (vsi Euroimmun, Lübeck, Nemčija) ter West Nile Virus IgG DxSelectTM ELISA in West Nile Virus IgM Capture DxSelectTM ELISA (Focus Diagnostics, Kalifornija, ZDA). Vzorce smo nato analizirali tudi s kemiluminescenčnimi monotesti VirClia® (Vircell, Granada, Španija) za KME, WNV in DENV. Pri VirClia® za KME smo ugotovili 95 % ujemanje z referenčnim testom za protitelesa IgG in 82,9 % ujemanje za protitelesa IgM. Pri VirClia® za DENV je bilo ujemanje 96,3 % pri protitelesih IgG in 85 % pri protitelesih IgM. Pri vzorcih bolnikov z WNV je bilo ujemanje med testi 100 %. Ugotovili smo, da se pri KMEV in DENV IgG ter WNV referenčni in preizkušani test odlično ujemata. Prav tako smo ugotovili tudi močno navzkrižno reaktivnost IgG med flavivirusi, zlasti med DENV in KMEV/WNV, kar lahko omeji uporabnost testov in zahteva previdno interpretacijo rezultatov, zlasti pri mejnih vrednostih. Arbovirus infections in humans can cause a wide range of symptoms, from asymptomatic infections to life-threatening conditions. Due to the short viremia in arbovirus infections, we often miss the diagnostic window for direct detection of the virus, but we can confirm the infection by detecting specific antibodies using various serological methods. The main limitation of serological methods is the high cross- reactivity between flaviviruses, which often makes it difficult to accurately identify the specific pathogen. This has prompted the development of new serological tools, such as chemiluminescence tests. We included 108 samples from patients with tick-borne meningoencephalitis, dengue, and West Nile fever, as well as 12 samples from healthy individuals in our master's thesis. To detect individual flavivirus infections, we used reference laboratory tests: Anti-TBE Virus ELISA IgM & IgG, Anti-Dengue Virus ELISA IgM & IgG (all Euroimmun, Lübeck, Germany), West Nile Virus IgG DxSelectTM ELISA and West Nile Virus IgM Capture DxSelectTM ELISA (Focus Diagnostics, California, USA). Samples were then analyzed using chemiluminescent monotests VirClia® (Vircell, Granada, Spain) for TBE, WNV and DENV. VirClia® for TBE showed 95 % agreement with the reference test for IgG antibodies and 82.9 % agreement for IgM antibodies. With VirClia® for DENV, the concordance was 96.3 % for IgG antibodies and 85 % for IgM antibodies. In samples from patients with WNV, the concordance between the tests was 100 %. We found that for TBEV and DENV IgG and for WNV, the reference test and the evaluated test concord perfectly. We also found strong cross-reactivity of IgG between flaviviruses, particularly between DENV and TBEV/WNV, which may limit the usefulness of the tests and require careful interpretation of the results, especially at borderline values. virusi flavivirusi standardne serološke metode kemiluminescenčni testi klinična opredelitev viruses flaviviruses standard serological methods chemiluminescence assay clinical evaluation true false false [P. Kuhar] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-04-07 10:19:11 2026-04-07 10:19:25 2026-04-08 04:27:28 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 579.61:616-078:578.833.2 279298 274375171 56879.pdf 56879.pdf 1 2D345D6BE78EEF2DE9C2174EED11E5F5 83eb3b0276fb108e9e62915d99ee69a83b91bddd75bdc58c800b05954836e5b5 4dafca0d-325a-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=231841 Biotehniška fakulteta 0 0 0