CATCH-EVseq, an Ultra-Sensitive Capture Assay to Monitor Human Enterovirus RNA
- 1Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA
Abstract
The detection of low-abundance viral RNA in chronic or persistent infections remains a major challenge in virology, constrained by the sensitivity and specificity of traditional methods. One such infection, chronic enterovirus (EV) infection, has been proposed as a potential underlying factor in Myalgic Encephalomyelitis (ME), although this hypothesis remains debated. Supporting evidence for EVs as a viral trigger includes historical spatiotemporal overlaps between ME outbreaks and poliomyelitis epidemics, frequent reports of “flu-like” illness at disease onset, seasonal outbreaks patterns, and serological findings. To address the problem of EV detection, we developed CATCH-EVseq, an ultra-sensitive platform built on the CATCH framework. By integrating high-throughput sequencing with cDNA capture using 49,847 probes targeting 3410 EV genomes, CATCH-EVseq achieved a detection limit in artificially created samples as low as one copy in a microgram of RNA for two out of three biological replicates. Additionally, spike-in controls allow for rigorous validation of the capture and sequencing throughout the CATCH-EVseq procedure. When applied to a small number of blood samples from both ME patients and healthy controls, CATCH-EVseq detected no EV RNA, either in direct assays or after in vitro viral amplification. While these results do not support the presence of circulating EV RNA in the blood of individuals with ME, they do not rule out persistent infection in other tissues or organs. Critically, CATCH-EVseq establishes a robust, ultra-sensitive framework for future investigations of low-level EV persistence in biopsies, post-mortem tissues, or other clinical contexts where chronic enterovirus infection is suspected.
Related
- Surveillance of Athletes’ Health in Resource-Constrained African Sports Contexts: Why Context Matters and Time To Catch Up
- From non-uptake during early adolescence to acceptance during a catch-up campaign: a qualitative study of changing HPV vaccination decisions across two life stages among young women in Amsterdam, the Netherlands
- Enterovirus 2B protein: From viroporin to membrane stress sensor, mechanistic principles of multi-pathway regulation.
- Beyond polio: advanced human in vitro models for studying non-polio enterovirus pathogenesis
- Invasive Enterovirus and Parechovirus Infections in Adults, Alberta, Canada, 2023–2025
BibTeX
@article{germain2026catchevseq,
title = {{CATCH-EVseq, an Ultra-Sensitive Capture Assay to Monitor Human Enterovirus RNA}},
author = {Arnaud Germain and Maureen R. Hanson},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = oct,
doi = {10.3390/ijms27209065},
url = {https://www.mdpi.com/1422-0067/27/20/9065},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - CATCH-EVseq, an Ultra-Sensitive Capture Assay to Monitor Human Enterovirus RNA
AU - Arnaud Germain
AU - Maureen R. Hanson
PY - 2026
DA - 2026/10/13
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27209065
UR - https://www.mdpi.com/1422-0067/27/20/9065
AB - The detection of low-abundance viral RNA in chronic or persistent infections remains a major challenge in virology, constrained by the sensitivity and specificity of traditional methods. One such infection, chronic enterovirus (EV) infection, has been proposed as a potential underlying factor in Myalgic Encephalomyelitis (ME), although this hypothesis remains debated. Supporting evidence for EVs as a viral trigger includes historical spatiotemporal overlaps between ME outbreaks and poliomyelitis epidemics, frequent reports of “flu-like” illness at disease onset, seasonal outbreaks patterns, and serological findings. To address the problem of EV detection, we developed CATCH-EVseq, an ultra-sensitive platform built on the CATCH framework. By integrating high-throughput sequencing with cDNA capture using 49,847 probes targeting 3410 EV genomes, CATCH-EVseq achieved a detection limit in artificially created samples as low as one copy in a microgram of RNA for two out of three biological replicates. Additionally, spike-in controls allow for rigorous validation of the capture and sequencing throughout the CATCH-EVseq procedure. When applied to a small number of blood samples from both ME patients and healthy controls, CATCH-EVseq detected no EV RNA, either in direct assays or after in vitro viral amplification. While these results do not support the presence of circulating EV RNA in the blood of individuals with ME, they do not rule out persistent infection in other tissues or organs. Critically, CATCH-EVseq establishes a robust, ultra-sensitive framework for future investigations of low-level EV persistence in biopsies, post-mortem tissues, or other clinical contexts where chronic enterovirus infection is suspected.
SN - 1422-0067
ER -