Detection of the Liman tick virus (unclassified <i>Chuviridae</i>) in tick cell line HAE/CTVM8
2025
Alexander G. Litov | Alexey M. Shchetinin | Ivan S. Kholodilov | Oxana A. Belova | Аnna S. Kalyanova | Vladimir A. Gushchin | Galina G. Karganova
Introduction. Tick cell lines are widely used to study the biology of ticks and tick-borne pathogens, especially viruses. Most of the cell cultures currently available have been obtained from tick embryonic cells and can be infected with viruses. The HAE/CTVM8 cell line was obtained from Hyalomma anatolicum ticks and is often used for isolation of novel viruses. The aim of the work is to study the HAE/CTVM8 cell line using high-throughput sequencing in order to search for viruses in it. Materials and methods. The HAE/CTVM8 cell culture fluid was ultracentrifuged. The resulting pellet was used for high-throughput sequencing after RNA extraction, reverse transcription reaction, and synthesis of the second strand. The resulting reads were filtered by length and quality in the Trimmomatic program, after which the contigs were assembled using the SPAdes program and analyzed for the presence of viral sequences. The final assembly of the virus genome was carried out in the Ugene program. Sequence alignment was performed by the MAFFT program. The phylogenetic trees were constructed using the IQ-TREE program. Results. We have identified the persistence of one virus, Liman tick virus (LMTV), in HAE/CTVM8 cell culture. Phylogenetically LMTV belongs to the Chuviridae – novel family, that consists of viruses detected by high-throughput sequencing, the virological characteristics of which are currently unknown. Conclusion. The obtained information is of significant importance when utilizing HAE/CTVM8 cell culture in scientific research and during the process of isolating new viruses. Our study shows that this cell line with persistent LMTV is a ready-to-use system for studying Chuviridae reproduction
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