Title Recovery of 5' and 3' utr sequences of wuhan mosquito virus 6
Translation of Title 5' ir 3' nekoduojančių Uhano uodų viruso 6 sekų nustatymas.
Authors Kazlauskaitė, Miglė
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Pages 68
Keywords [eng] UTR, untranslated region, WuMV-6, Wuhan mosquito virus 6, RACE, rapid amplification of copy DNA ends, RNA, hyp1, hypothetical segment 1, NP, nucleoprotein, orthomyxovirus.
Abstract [eng] Orthomyxoviruses represent a diverse family of segmented RNA viruses including major human and animal pathogens such as influenza viruses. Recent metagenomic studies have revealed numerous arthropod-associated orthomyxoviruses, including Wuhan Mosquito Virus 6 (WuMV-6), which has achieved exceptional global distribution across six continents. Understanding viral biology requires characterization of untranslated regions (UTRs) that serve critical regulatory functions. Yet UTR sequences remain completely unknown for WuMV-6 and related quaranjaviruses, preventing development of essential molecular tools. This study aimed to characterize the 5′ and 3′ UTRs of all eight WuMV-6 genome segments using samples from Lithuanian Culex pipiens mosquito populations. Reverse transcription quantitative polymerase chain reaction screening identified WuMV-6-positive pools. Those were later subjected to rapid amplification of complementary DNA ends (RACE) followed by Oxford Nanopore sequencing. WuMV-6 was detected in 42% of mosquito pools across 12 of 25 sampling localities with an estimated individual prevalence of 1,45%, confirming widespread circulation throughout Lithuania. Phylogenetic analysis revealed that recovered sequences clustered within the Baltic/Northern European lineage, distinct from the global lineage. However, RACE analysis encountered significant technical challenges including non-specific amplification and low recovery rates. Only four authentic WuMV-6 sequences were recovered from 57 samples, representing fragments from hypothetical segment 1 and nucleoprotein segments with insufficient sequencing depth for reliable analysis. Complete UTR characterization was not achieved due to methodological limitations. The study demonstrates the need for improved approaches to characterize UTRs in newly discovered arthropod-associated orthomyxoviruses, as current techniques prove insufficient for these viral lineages.
Dissertation Institution Vilniaus universitetas.
Type Master thesis
Language English
Publication date 2026