Non-Targeted RNA Sequencing: Towards the Development of Universal Clinical Diagnosis Methods for Human and Veterinary Infectious Diseases

• Published in: Veterinary sciences Vol. 11; no. 6; p. 239
• Main Authors: Spatz, Stephen, Afonso, Claudio L
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 26-05-2024; MDPI
• Subjects: Automation; Bacteria; Bioinformatics; clinical diagnostics; Communicable diseases; Diagnosis; Environmental monitoring; Fungi; Gene expression; Genetic engineering; Genomes; Infectious diseases; International organizations; Medicinal plants; Metagenomics; Methods; next-generation sequencing; NGS; non-targeted; Pathogens; Plant diseases; Review; Ribonucleic acid; RNA; RNA sequencing; Veterinary medicine; Viruses; agnostic diagnostics; RNA; clinical diagnostics; next-generation sequencing; metagenomics; veterinary; NGS; human; non-targeted
• ISSN: 2306-7381; 2306-7381
• DOI: 10.3390/vetsci11060239

Metagenomics offers the potential to replace and simplify classical methods used in the clinical diagnosis of human and veterinary infectious diseases. Metagenomics boasts a high pathogen discovery rate and high specificity, advantages absent in most classical approaches. However, its widespread adoption in clinical settings is still pending, with a slow transition from research to routine use. While longer turnaround times and higher costs were once concerns, these issues are currently being addressed by automation, better chemistries, improved sequencing platforms, better databases, and automated bioinformatics analysis. However, many technical options and steps, each producing highly variable outcomes, have reduced the technology's operational value, discouraging its implementation in diagnostic labs. We present a case for utilizing non-targeted RNA sequencing (NT-RNA-seq) as an ideal metagenomics method for the detection of infectious disease-causing agents in humans and animals. Additionally, to create operational value, we propose to identify best practices for the "core" of steps that are invariably shared among many human and veterinary protocols. Reference materials, sequencing procedures, and bioinformatics standards should accelerate the validation processes necessary for the widespread adoption of this technology. Best practices could be determined through "implementation research" by a consortium of interested institutions working on common samples.