Insights into the Sequencing of Viral Pathogens associated with Acute Respiratory Infection

The respiratory tract may contain many varieties of commensal and pathogenic organisms based on the host environment. Detection of a species alone may not present the most complete view of the biological analysis of a sample.  The Twist Respiratory Virus Research Panel enables the enrichment and sequencing of respiratory viral pathogens, allowing researchers to detect the species and obtain sequence information about the viral genomes.

Additionally, acute respiratory infection symptoms can overlap with those from other respiratory viruses. The Twist Respiratory Virus Research Panel can help researchers identify the cause of infection and identify co-infection.

KEY BENEFITS

Comprehensive Detection of Respiratory Pathogens

  • Designed to target multiple viruses in a single capture
  • Targeted against the reference sequences for 29 common human respiratory viruses

Reliable Detection of Co-Infections

  • Determine if a sample contains multiple pathogens
  • Add additional pathogens by customizing the panel

Highly Sensitive at Low Viral Loads

  • Gather data from many sample types including nasopharyngeal samples
  • Detect viral loads as low as 100 viral genome copies

This panel's design incorporates the use of the CATCH (Compact Aggregation of Targets for Comprehensive Hybridization) algorithm. For additional details on CATCH, please refer to Metsky HC and Siddle KJ et al. Capturing sequence diversity in metagenomes with comprehensive and scalable probe design. Nature Biotechnology, 37(2), 160–168 (2019). doi: 10.1038/ s41587-018-0006-x

NGS Target Enrichment of Viral Pathogens

Detect Common Human Respiratory Pathogens with a Single Test

Compiled from the GenBank database, the 41,047 probes in this panel will allow researchers to separate COVID-19 symptoms from those of other respiratory illnesses, both influenza- and non-influenza-related. The Twist Respiratory Panel enables enrichment of these viral sequences for high-resolution NGS. This leads to high-sensitivity detection of as little as 100 copies of viral material with more than 5000x enrichment even from challenging samples, and coverage of > 99.9% of the genome at 1X or greater post-enrichment.

Detection of viral standards from different viral families

Each Twist Synthetic Viral Control was spiked into 50 ng of human carrier RNA at 1,000,000 copies prior to cDNA synthesis. For each standard, we show percent of on-target viral reads, fold enrichment above input, median coverage across the genome, the percent of bases with at least 30x coverage, and the template length. Viral genomes are ordered by template length (smallest to longest) among all genome segments. In most cases, we found that over 70 percent of reads came from viral genomes in these libraries, representing at least a 2500-fold enrichment over the spiked-in content.

Multiplex for Higher Throughput and Lower Cost

A comparison of the percent on-target viral reads captured using the Twist Synthetic Viral Controls at different viral titers, using a multiplex and single-plex hybridization reaction. The first graph shows comparisons using a multiplex and single-plex hybridization reaction using 1,000,000 copies per library, while the second graph shows a similar comparison using 100, 10,000, and 1,000,000 copies per library. Each viral control was captured using the Twist Standard Target Enrichment workflow and the Twist Respiratory Virus Research Panel. This data demonstrates that 8-plex capture gives comparable efficient enrichment as a single plex capture when using the Twist Respiratory Virus Research Panel.

One Codex Analysis

Twist Respiratory Virus Research Panel comes with access to the One Codex cloud-based tool. Ideal for research and clinical use, as it can be used to create publication-ready visuals and printable reports for labs preparing laboratory developed tests.

When using the Twist Respiratory Virus Research Panel, the One Codex analysis platform is accessible for all customers. This platform is ideal for both research and clinical use cases, including production of publication-ready visuals, shareable reports, and other analysis tools. When purchasing the panel you will be provided with onboarding instructions and credits for sample analysis on the One Codex platform. Learn more about this microbial sample analysis solution at One Codex.

Twist Respiratory Virus Research Panel
Twist Respiratory Virus Research Controls

NGS Target Enrichment of Viral Pathogens

Twist Total Nucleic Acids Library Preparation Kit
Twist Target Enrichment Protocol
Twist Fast Hybridization Target Enrichment Protocol