20.500.12556/RUL-190034 Primerjava metod za izolacijo bakteriofagov iz različnih naravnih okolij Testing different methods for isolating bacteriophages from different natural environments Procesi za izolacijo bakteriofagov iz okoljskih vzorcev so ključen predmet raziskav za učinkovito iskanje bakteriofagov v okolju in predstavljajo glavni način identifikacije novih bakteriofagov, ki so zmožni inficirati na antibiotike odporne patogene bakterije. Ti so za medicino zelo relevantni kot alternativa uporabi antibiotikov, ki so za boj proti okužbam nekaterih bakterijskih vrst neučinkoviti, zaradi njihove vsesplošne razširjenosti, pa jih je moč najti skoraj kjerkoli v okolju. Za izolacijo iz okoljskih vzorcev so bile razvite številne metode, štiri so bile preizkušene in primerjane v sklopu te naloge. Izmed preizkušenih postopkov je najvišji izkoristek fagov v osnovnem vzorcu dosegla anionska izmenjevalna kromatografija, a z najvišjimi finančnimi stroški materiala in opreme ter najvišjo zahtevnostjo izvedbe. Šaržna kromatografija s hidroksiapatitom predstavlja cenovno bolj ugodno in enostavnejšo alternativo s primerljivim izkoristkom, a z višjo standardno napako, medtem ko precipitacija s polietilen glikolom predstavlja še cenejšo in do fagnih delcev bolj nežno metodo, ki pa trpi za slabimi izkoristki. Metoda z obogatitvijo fagne suspenzije ni bila uspešno izvedena z izbranim postopkom, zaradi česar se te metode ni dalo primerjati z ostalimi. Optimalna izbira metode za izolacijo bakteriofagov iz okoljskih vzorcev je po ugotovitvah v tem delu torej odvisna od potreb in vložka izvajalca. The processes for bacteriophage isolation are a key subject of study for efficient phage hunting, which represents the main way of discovery of novel bacteriophages capable of infection antibiotic resistant pathogenic bacteria. These are highly medically relevant as an alternative to antibiotics, which are inefficient in combating certain infections, and are able to be found almost anywhere in the envirnment due to their ubiquity. Numerous methods for bacteriophage isolation have been developed, four of which have been tested and compared in the scope of this work. Of the tested procedures, anion exchange chromatography yielded the highest recovery, although with also the highest financial cost per processed sample and the highest level of complexity of execution. Batch chromatography using hydroxyapatite represents a cheaper and simpler alternative with comparable recovery but a significantly larger standard error, while precipitation with polyethylene gylcol remains an even cheaper alternative, which suffers from a significantly worse recovery. The phage suspension enrichment method was not performed successfully with the chosen procedure, hence it wasn't compared with the rest of the methods. The optimal choice of method for phage hunting, as indicated by the findings of this work, is therefore subject to the needs and resources of the phage hunter. bakteriofagi fagi fagna terapija izolacija okoljski vzorci precipitacija kromatografija Bakteriophages phages phage therapy isolation environmental samples precipitation chromatography true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-10-21 07:16:08 2026-10-21 07:16:16 2026-10-23 10:37:16 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 299387 302219011 64865.pdf 64865.pdf 1 E727571720E989106E583040C3B11EB8 7482c1c66ca1834b90001782ff563070f20a5545a62eb571c1c69ab7f4220b19 61dd1b60-cd0e-11f1-8bc5-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=251023 Biotehniška fakulteta 0 0 0