20.500.12556/DiRROS-27862 Izzivi uporabe visoko-pretočnega sekvenciranja (HTS) za rutinsko diagnostiko viroidov Challenges in the application of high-throughput sequencing for routine diagnostics of viroids Viroidne bolezni predstavljajo nenehno grožnjo svetovnemu kmetijstvu, saj lahko ti majhni in strukturno preprosti patogeni okužijo širok spekter gospodarsko pomembnih rastlin. Učinkovito preprečevanje in obvladovanje viroidnih okužb zato zahteva diagnostične pristope, ki omogočajo zanesljivo, občutljivo in specifično identifikacijo povzročiteljev bolezni. Z razvojem biokemije in molekularne biologije se je v zadnjih letih izboljšala občutljivost in specifičnost številnih diagnostičnih metod. Z zniževanjem stroškov sekvenciranja in večjo dostopnostjo se zelo hitro povečuje zanimanje za uporabo tehnologij visokopretočnega sekvenciranja (HTS) v rutinski rastlinski diagnostiki. Kljub velikemu potencialu HTS pa uvedba teh tehnologij v laboratorijsko prakso ostaja zahtevna. Med ključne izzive sodijo izbira ustrezne sekvenčne platforme in laboratorijskih protokolov, uporaba primernih bioinformacijskih orodij, zagotavljanje strokovno usposobljenega kadra ter vzpostavitev ustrezne računalniške infrastrukture. Čeprav so že oblikovani določeni standardi in smernice za uporabo HTS, ti zaradi specifičnih potreb laboratorijev niso univerzalno uporabni. Pregledni članek obravnava ključne vidike različnih pristopov HTS, njihove prednosti in omejitve ter prikazuje njihovo uporabnost na primeru diagnostike viroidnih bolezni. Viroid diseases pose a continual threat to global agriculture, as these small and structurally simple pathogens are capable of infecting a wide range of economically important plant species. Effective prevention and management of viroid infections therefore require diagnostic approaches that ensure reliable, sensitive, and specific identification of the causal agents. Advances in biochemistry and molecular biology have, in recent years, significantly improved the sensitivity and specificity of many diagnostic methods. With decreasing sequencing costs and increasing accessibility, interest in the use of high-throughput sequencing (HTS) in routine plant diagnostics has grown rapidly. Despite the great potential of HTS, the implementation of these technologies into laboratory practice remains challenging. Key obstacles include the selection of an appropriate sequencing platform and laboratory protocols, the use of suitable bioinformatic tools, the availability of adequately trained personnel, and the establishment of suitable computational infrastructure. Although certain standards and guidelines for HTS have already been developed, they are not universally applicable due to the specific needs of individual laboratories. This review article discusses the key aspects of different HTS approaches, their advantages and limitations, and illustrates their applicability using the example of viroid disease diagnostics. diagnostika visoko-pretočno sekvenciranje (HTS) nanopore tehnologija (ONT) bioinformatika viroidi diagnostics high-throughput sequencing (HTS) Oxford Nanopore technology (ONT) bioinformatics viroids true false true Slovenski jezik Angleški jezik Inštitut za hmeljarstvo in pivovarstvo Slovenije Neznano 2026-02-26 12:02:29 2026-02-26 12:02:29 2026-03-07 03:51:38 0000-00-00 00:00:00 2025 0 0 Nasl. z nasl. zaslona; Opis vira z dne 19. 12. 2025; str. 72-84 Št 32 2025 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 2025-12-19 632.38 2536-1988 262352387 https://www.dlib.si/details/URN:NBN:SI:spr-OS4I5CDJ 1 a55b87f8-1302-11f1-9cb0-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=40997 IZZIVI_UPORABE_VISOKO-PRETOCNEGA_SEKVENCIRANJA_(HTS)_ZA_RUTINSKO.pdf IZZIVI_UPORABE_VISOKO-PRETOCNEGA_SEKVENCIRANJA_(HTS)_ZA_RUTINSKO.pdf 1 C1A4C45B8E179916AFE5A1A4940FE00A 90329a9c9de4860106df22fdbe20fcd8137f4a3df83c1829896734af1c0f969f 8cd80c40-1947-11f1-81cd-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=41357 Inštitut za hmeljarstvo in pivovarstvo Slovenije Hmeljarski bilten 0 0 0