PREreviews of “TITAN-RNA: A hybrid-capture sequencing panel detects known and unknown Flaviviridae for diagnostics and vector surveillance”

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TITAN-RNA: A hybrid-capture sequencing panel detects known and unknown Flaviviridae for diagnostics and vector surveillance

by Chloe M. Murrell, Aiman Sabaawy, Xiyu Wang, Nagi Hashimoto, Kristina Ceres, Yining Sun, Jordan D. Zehr, Aine Lehane, Emily Mader, Natalie Bailey, Marie V. Lilly, Laura Plimpton, Guillaume Reboul, Joel J. Brown, Kelly Sams, Lauren Singh, Ethan Seiz, Ellie Bourgikos, Chantal B.F. Vogels, Alexander T. Ciota, Victoria Schnurr, Jennifer Grenier, Xavier Berthet, Ana I. Bento, Laura Harrington, Maria Diuk-Wasser, Ximena Olarte-Castillo, and Laura B. Goodman

Posted: June 8, 2025
Server: bioRxiv
DOI: 10.1101/2025.06.08.658352

Abstract

Clinical testing and public health surveillance can be significantly improved by incorporating sequencing-based molecular detection and subtyping for real-time monitoring of virus evolution. With phylogenetic analysis used for speciation and variant subtyping, target analyte specificity can be relaxed well beyond typical parameters acceptable in PCR-based diagnostics. Hybrid capture is a promising way to enrich large numbers of sequences with maximal flexibility, using standard molecular biology laboratory equipment and small benchtop sequencers. Here, we report the development and bench validation of a hybrid capture based next-generation sequencing diagnostic panel for all known viral tick-borne pathogens, TITAN-RNA. Based on systematic testing with simulated novel viruses and field samples, we determined a 10% tolerance for evenly distributed mutations or 27% tolerance for naturally occurring viral divergence. The TITAN-RNA extrapolated limit of detection in blood is 19.1 genome copies by complementary log-log analysis, and linearity performance (R ² ≥ 0.99) is amenable for its use as a quantitative assay. As proof of principle for public health surveillance and evolutionary studies, we report two putatively novel segmented Flavi- like viruses in New York State, USA, identified from the invasive Haemaphysalis longicornis tick.

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PREreview by Ananddeep Patel

Authored by Ananddeep Patel

This preprint describes TITAN-RNA, a hybrid-capture next-generation sequencing panel designed to detect known and potentially divergent or previously uncharacterized Flaviviridae and other tick-borne viral pathogens for diagnostic and vector-surveillance applications. The study reports bench…

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PREreviews of TITAN-RNA: A hybrid-capture sequencing panel detects known and unknown Flaviviridae for diagnostics and vector surveillance

1 PREreview