Abstract
Viruses are evolving at an alarming rate, spreading and inconspicuously adapting to cutting-edge therapies. Therefore, the search for rapid, informative and reliable diagnostic methods is becoming urgent as ever. Conventional clinical tests (PCR, serology, etc.) are being continually optimized, yet provide very limited data. Could high throughput sequencing (HTS) become the future gold standard in molecular diagnostics of viral infections? Compared to conventional clinical tests, HTS is universal and more precise at profiling pathogens. Nevertheless, it has not yet been widely accepted as a diagnostic tool, owing primarily to its high cost and the complexity of sample preparation and data analysis. Those obstacles must be tackled to integrate HTS into daily clinical practice. For this, three objectives are to be achieved: (1) designing and assessing universal protocols for library preparation, (2) assembling purpose-specific pipelines, and (3) building computational infrastructure to suit the needs and financial abilities of modern healthcare centers. Data harvested with HTS could not only augment diagnostics and help to choose the correct therapy, but also facilitate research in epidemiology, genetics and virology. This information, in turn, could significantly aid clinicians in battling viral infections.
Highlights
As some statistical models suggest, there are over 320,000 mammalian viruses in existence [1], a little over 200 of which are known to infect humans [2], with the number increasing steadily over decades [3]
Various approaches exist that feature Whole genome sequencing (WGS), amplicon sequencing, enrichment sequencing and metagenomics, the choice depending on the type of pathogen and experiment objective
Metagenomics, for instance, works best with unknown viruses, whereas PCR-based techniques are apt for samples with low diversity and short genomes
Summary
As some statistical models suggest, there are over 320,000 mammalian viruses in existence [1], a little over 200 of which are known to infect humans [2], with the number increasing steadily over decades [3]. We are attempting to look at a vast sea of threats through the eye of a needle As of this day, a multitude of clinical tests are available for detection of viruses, e.g., FISH, ELISA (enzyme-linked immunosorbent assay), and PCR- and their numerous modifications. With the entirety of human-infecting viruses, most of which are yet to be revealed, it becomes increasingly difficult to maintain up-to-date primer panels while ensuring the correct reaction conditions and high specificity for differentiating closely related viral species This situation is aggravated by the low rates of correct pathogen identification [9]. We hope to demonstrate how modern clinical laboratories would benefit from adopting HTS as a routine diagnostic technique
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