Overview
TruSight Oncology 500 ctDNA v2 is a pan-cancer NGS assay that enables in-house comprehensive genomic profiling (CGP) of circulating tumor DNA (ctDNA) in blood plasma for research.
Target 523 genes to assess DNA variants across major variant classes (SNV, MNV, indels, CNV, and gene rearrangements), plus key immuno-oncology genomic signatures (bTMB, bMSI).
Streamline workflow with advanced chemistry and automation-friendly kits to enable a single-day library prep plus enrichment and total turnaround time of 2.5-4 days.
Leverage low cfDNA inputs (20 ng) from minimally invasive blood samples to achieve sensitive and accurate detection (0.2% VAF for SNVs) using UMI-based hybrid-capture library prep and deep sequencing.
Take advantage of transformative economics, fast sequencing run times, and broad batch sizes by implementing the assay on NovaSeq X Series (4-128 samples) or NextSeq 2000 System.
Use DRAGEN secondary analysis for rapid variant calling on-premises or in the cloud with Illumina Connected Analytics. Access insights using integrated reporting with Illumina Connected Insights or Velsera.
Generate accurate variant results with fragmentomics-and frequency-based identification of tumor, germline, and clonal hematopoiesis variants. Leverage Maximum Somatic Allele Frequency (MSAF) to estimate tumor fraction.
Enable CGP with a comprehensive portfolio
Maximize chances of identifying relevant alterations with the TruSight Oncology 500 product line.
TruSight Oncology 500 v2
Enable CGP findings faster from FFPE tissue with a more efficient workflow, using a comprehensive pan-cancer panel that covers all major variant classes and genomic signatures (TMB, MSI, and HRD).
*TruSight Oncology v2 is not available for sale in Japan.
Example workflow
TruSight Oncology 500 ctDNA v2 Assay
NextSeq 2000 Systems
NovaSeq 6000 System
NovaSeq 6000Dx (in RUO mode) *
NovaSeq X Series
DRAGEN secondary analysis: on-premise via DRAGEN server
DRAGEN secondary analysis: cloud-based via Illumina Connected Analytics (ICA)
Project recommendations
Instrument: Recommended number of samples
NextSeq 2000 System: 4 samples per run (P4 flow cell), 800M paired-end reads, 35,000* coverage
4 samples per run (P4 flow cell), 800M paired-end reads, 35,000* coverage
2 × 150 bp
NovaSeq 6000 System: 4 samples per run (S1 flow cell), 800M paired-end reads, 35,000x coverage
4 samples per run (S1 flow cell), 800M paired-end reads, 35,000x coverage
2 x 150 bp
NovaSeq 6000 System: 8 samples per run (S2 flow cell), 800M paired-end reads, 35,000x coverage
8 samples per run (S2 flow cell), 800M paired-end reads, 35,000x coverage
NovaSeq 6000 System: 24 samples per run (S4 flow cell), 800M paired-end reads, 35,000x coverage
24 samples per run (S4 flow cell), 800M paired-end reads, 35,000x coverage
NovaSeq X System: 4 samples per run (1.5B flow cell), 800M paired-end reads, 35,000x coverage
4 samples per run (1.5B flow cell), 800M paired-end reads, 35,000x coverage
NovaSeq X System: 64 samples per run (25B flow cell), 800M paired-end reads, 35,000x coverage
64 samples per run (25B flow cell), 800M paired-end reads, 35,000x coverage
NovaSeq X System: 24 samples per run (10B flow cell), 800M paired-end reads, 35,000x coverage
24 samples per run (10B flow cell), 800M paired-end reads, 35,000x coverage
Related applications and methods
NGS in oncology
Next-generation sequencing provides deep insights into the molecular underpinnings of tumors and can help advance the promise of personalized medicine.
Immuno-oncology research
Next-generation sequencing enables immuno-oncology researchers to gain insights into immunotherapy response factors and tumor immune evasion mechanisms.
Pathology and clinical cancer research
Our clinical cancer research solutions deliver accurate genomic information, and enable labs to analyze multiple genes in a single test.
SNV detection at low VAF
Synthetic control samples with known variant allele frequency (VAF) for each single nucleotide variant (SNV) were diluted to values ranging from 0.20%–0.50% VAF and analyzed by TruSight Oncology 500 ctDNA v2.
Indel detection at low VAF
Synthetic control samples with known VAF for each insertion or deletion were diluted to values ranging from 0.20%–0.50% VAF and analyzed by TruSight Oncology 500 ctDNA v2.
High TMB correlation compared to tumor-normal calling
Tumor-only workflow of TSO 500 ctDNA v2 utilizing advanced bioinformatics for germline and CH variant filtering produces highly concordant bTMB compared to a tumor-normal workflow.
Sensitive MSI detection for immuno-oncology research
Microsatellite instability (MSI) was evaluated in 3 cell lines with known MSI-high status (samples 1–3) and detected down to 0.3% tumor fraction assessing up to ~2300 homopolymer sites.
Analytical performance for CNVs, LOD ≥1.3 fold for amplifications and ≤0.6 for deletions
Lower relative MYC detection due to limit of detection approached faster due to fewer starting copies. Illumina data on file, 2023.
Analytical performance for gene rearrangements with detection at LOD (0.5%)
VAF calculated by dividing the total number of supporting reads by the higher depth of the two sides of the breakpoint. Illumina data on file, 2023.
TruSight Oncology 500 ctDNA v2 video
Illumina improved the analysis of circulating tumors with TruSight Oncology ctDNA 500 v2. Learn about new features and benefits in the video.
TruSight Oncology 500 portfolio brochure
Learn more about the TruSight Oncology 500 portfolio of products.
Illumina collaborates with National Cancer Center Japan to address a leading cause of death in Asia
Learn about this joint effort to accelerate the development of personalized treatments based on genomic information.
TruSight Oncology 500 ctDNA v2 (24 samples)
20105899
TSO 500 ctDNA v2 Manual Kit View less
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TruSight Oncology 500 ctDNA v2 for Use with NovaSeq 6000 S2 Flow Cell (24 samples)
20105901
Mixed Workflow TSO 500 ctDNA v2 + S2 FC Manual Kit View less
TruSight Oncology 500 ctDNA v2 for Use with NovaSeq 6000 S4 Flow Cell (24 samples)
20105902
Mixed Workflow TSO 500 ctDNA v2 + S4 FC Manual Kit View less
TruSight Oncology 500 ctDNA v2 (24 samples) plus Velsera Interpretation Report
20105905
TSO 500 ctDNA v2 Manual Kit + Velsera Interpretation Report View less
TruSight Oncology 500 ctDNA v2 plus Velsera Interpretation Report, for Use with NovaSeq 6000 S2 Flow Cell (24 samples)
20105907
Mixed Workflow TSO 500 ctDNA v2 + S2 FC Manual Kit + Velsera Interpretation Report View less
TruSight Oncology 500 ctDNA v2 plus Velsera Interpretation Report, for Use with NovaSeq 6000 S4 Flow Cell (24 samples)
20105908
Mixed Workflow TSO 500 ctDNA v2 + S4 FC Manual Kit + Velsera Interpretation Report View less
TruSight Oncology 500 ctDNA Kit (48 samples)
20039252
TruSight Oncology 500 ctDNA Kit (Includes DNA library prep and enrichment reagents. Does not include NovaSeq core reagents) View less
TruSight Oncology 500 ctDNA Kit plus Velsera Interpretation Report (16 indexes, 48 Samples)
20043410
Includes library prep and enrichment reagents, plus data interpretation reports (through Velsera Clinical Genomics Workspace software). Does not include NovaSeq System Core Reagents. View less
TruSight Oncology 500 ctDNA Kit, for use with NovaSeq 6000 S2 (48 samples)
20101995
Combination SKU for TSO500 ctDNA and the NovaSeq 6000 S2 Core consumables View less
TruSight Oncology 500 ctDNA Kit, for use with NovaSeq 6000 S4 (48 samples)
20101998
Combination SKU for TSO500 ctDNA and the NovaSeq 6000 S4 Core consumables View less
TruSight Oncology 500 ctDNA Kit for use with NovaSeq 6000 S2 plus Velsera interpretation report (16 indexes, 48 samples)
20102000
Combination SKU for TSO500 ctDNA and the NovaSeq 6000 S2 Core consumables + Velsera report View less
TruSight Oncology 500 ctDNA Kit for use with NovaSeq 6000 S4 plus Velsera interpretation report (16 indexes, 48 samples)
20102001
Combination SKU for TSO500 ctDNA and the NovaSeq 6000 S4 Core consumables + Velsera report View less
TruSight Oncology 500 ctDNA v2 (24 samples) plus Illumina Connected Insights Software
20105911
TSO 500 ctDNA v2 Manual Kit + Illumina Connected Insights Software View less
TruSight Oncology 500 ctDNA v2 plus Illumina Connected Insights Software, for Use with NovaSeq 6000 S2 Flow Cell (24 samples)
20105913
Mixed Workflow TSO 500 ctDNA v2 + S2 FC Manual Kit+ Illumina Connected Insights Software View less
TruSight Oncology 500 ctDNA v2 plus Illumina Connected Insights Software, for Use with NovaSeq 6000 S4 Flow Cell (24 samples)
20105914
TruSight Oncology 500 ctDNA v2 plus Illumina Connected Insights Software, for Use with NovaSeq 6000 S4 Flow Cell (24 samples) View less
IDT® for Illumina® UMI DNA/RNA UD Indexes Set A, Ligation (96 Indexes, 96 Samples)
20034701
UMI DNA Index Anchors (Plate = 20027219, Box = 20032119) + Nextera Compatible Unique Dual Index A (Sales Kit = 20027213, Plate = 20025019, Box = 20026121) View less
IDT® for Illumina® UMI DNA/RNA UD Indexes Set B, Ligation (96 Indexes, 96 Samples)
20034702
UMI DNA Index Anchors (Plate = 20027219, Box = 20032119) + Nextera Compatible Unique Dual Index B (Sales Kit = 20027214, Plate = 20025080, Box = 20026930) View less
IDT for Illumina UMI DNA/RNA Index Anchors Set A for Automation
20066404
Includes one box of 96 IDT for Illumina - UMI DNA/RNA UD Indexes Set A for Automation sufficient for labeling 96 samples and one box of 96 IDT for Illumina - UMI DNA Index Anchors for Automation. View less
IDT for Illumina UMI DNA/RNA Index Anchors Set B for Automation
20063213
Includes one box of 96 IDT for Illumina - UMI DNA/RNA UD Indexes Set B for Automation sufficient for labeling 96 samples and one box of 96 IDT for Illumina - UMI DNA Index Anchors for Automation. View less
NovaSeq 6000 S2 Reagent Kit v1.5 (300 cycles)
20028314
Includes one S2 flow cell, one buffer cartridge, one cluster cartridge, and one sequencing cartridge to support a 300-cycle run on the NovaSeq 6000 System. View less
NovaSeq 6000 S4 Reagent Kit v1.5 (300 cycles)
20028312
Includes one S4 flow cell, one buffer cartridge, one cluster cartridge, and one sequencing cartridge to support a 300-cycle run on the NovaSeq 6000 System. View less
NovaSeq™ X Series 1.5B Reagent Kit (300 Cyc)
20104705
This reagent kit provides one NovaSeq X Series 1.5B flow cell (with 2- lanes) and reagent consumables to support a single flow cell 300 cycles run on the NovaSeq X and NovaSeq X Plus systems. View less
NovaSeq X Series 10B Reagent Kit (300 Cycle)
20085594
This reagent kit provides one NovaSeq X Series 10B flow cell (with 8- lanes) and reagent consumables to support a single flow cell 300 cycles run on the NovaSeq X and NovaSeq X Plus systems. View less
Illumina DRAGEN Server v4
20051343
Includes Advance Exchange support for the first year. Requires purchase of annual DRAGEN license. View less
ICA Basic Annual Subscription
20044874
Illumina Connected Analytics (ICA) Basic Annual Subscription. This product includes 1 year of access to ICA Basic, including sequencing instrument connectivity, data management capabilities, and access to pre-packaged analysis tools. View less
ICA Professional Annual Subscription
20044876
Illumina Connected Analytics (ICA) Professional Annual Subscription. This product includes 1 year of access to ICA, including sequencing instrument connectivity, data management capabilities, access to pre- packaged tools, and the ability to create customized workflows composed of tools, pipelines, data warehouses, and notebooks. View less
ICA Enterprise Annual Subscription
20038994
Illumina Connected Analytics (ICA) Enterprise Annual Subscription. This product includes 1 year of access to ICA Enterprise, including sequencing instrument connectivity, data management capabilities, custom and pre-packaged analysis tools, and the Base module for data warehousing and mining. ICA Enterprise also includes optional HIPAA BAA (US-only), single sign-on (SSO), and a service level agreement (SLA). View less
ICA Enterprise Srvc & Compliance Add-on
20066830
Illumina Connected Analytics (ICA) Compliance enables single sign-on (SSO), multi-factor authentication (MFA), HIPAA BAA (US-only), and a Service Level Agreement (SLA) for an ICA Basic Annual Subscription. View less
Illumina Analytics - 1 iCredit
20042038
iCredits are used for data storage and analysis on either BaseSpace Sequence Hub or Illumina Connected Analytics. View less
Illumina Analytics Starter Pack - 1,000 iCredits
20042039
Illumina Analytics - 5,000 iCredits
20042040
Illumina Analytics - 50,000 iCredits
20042041
Illumina Analytics - 100,000 iCredits
20042042
Insights - Annual Subscription
20090137
Annual subscription to Illumina Connected Insights platform. View less
Illumina Connected Insights – Trial Subscription
20112516
Free trial subscription for Illumina Connected Insights - for oncology. View less
Illumina Connected Insights – Oncology Genome Equivalent Sample - VCF
20090138
Illumina Connected Insights pre-paid oncology samples on per genome equivalent basis starting from VCF: one genome is equivalent to 2 exomes, 3 large and 6 small panel samples. Any unused samples will automatically roll over provided that the annual subscription to Illumina Connected Insights (20090137) is renewed on an annual basis. Access to a set of oncology knowledge bases is included. View less
Connected Insights Training - Remote
20092376
Illumina Connected Insights Training - Remote includes five (5) hours of product training delivered virtually. View less
Informatics Professional Services - Remote
20071787
Professional services for Illumina informatics products and solutions including software implementations, pipelines and workflow development, integration, networking, migration, data ingestion projects and other informatics consultative, project-based services, performed remotely, and defined by a statement of work. View less
TruSight Oncology 500 ctDNA v2 for Automation (48 samples)
20105900
TSO 500 ctDNA v2 AutomatedKit View less
TruSight Oncology 500 ctDNA v2 Automation Kit, for Use with NovaSeq 6000 S2 Flow Cell (48 samples)
20105903
Mixed Workflow TSO 500 ctDNA v2 + S2 FC Automated Kit View less
TruSight Oncology 500 ctDNA v2 Automation Kit, for Use with NovaSeq 6000 S4 Flow Cell (48 samples)
20105904
Mixed Workflow TSO 500 ctDNA v2 + S4 FC Automated Kit View less
TruSight Oncology 500 ctDNA v2 for Automation (48 samples) plus Velsera Interpretation Report
20105906
TSO 500 ctDNA v2 AutomatedKit + Velsera Interpretation Report View less
TruSight Oncology 500 ctDNA v2 Automation Kit plus Velsera Interpretation Report, for Use with NovaSeq 6000 S2 Flow Cell (48 samples)
20105909
Mixed Workflow TSO 500 ctDNA v2 + S2 FC Automated Kit + Velsera Interpretation Report View less
TruSight Oncology 500 ctDNA v2 Automation Kit plus Velsera Interpretation Report, for Use with NovaSeq 6000 S4 Flow Cell (48 samples)
20105910
Mixed Workflow TSO 500 ctDNA v2 + S4 FC Automated Kit + Velsera Interpretation Report View less
TruSight Oncology 500 ctDNA v2 for Automation (48 samples) plus Illumina Connected Insights Software
20105912
TSO 500 ctDNA v2 AutomatedKit+ Illumina Connected Insights Software View less
TruSight Oncology 500 ctDNA v2 Automation Kit plus Illumina Connected Insights Software, for Use with NovaSeq 6000 S2 Flow Cell (48 samples)
20105915
Mixed Workflow TSO 500 ctDNA v2 + S2 FC Automated Kit+ Illumina Connected Insights Software View less
TruSight Oncology 500 ctDNA v2 Automation Kit plus Illumina Connected Insights Software, for Use with NovaSeq 6000 S4 Flow Cell (48 samples)
20105916
Mixed Workflow TSO 500 ctDNA v2 + S4 FC Automated Kit+ Illumina Connected Insights Software View less
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How does TruSight Oncology 500 ctDNA v2 differ from other TruSight Oncology assays?
TruSight Oncology 500 ctDNA v2 is an ultra-sensitive, streamlined liquid biopsy assay that enables CGP from blood plasma samples in 2.5-4 days. TruSight Oncology 500 ctDNA v2 offers improved sensitivity using a lower cfDNA input, a faster turnaround time, and a more streamlined workflow compared to the original TruSight Oncology 500 ctDNA assay. TruSight Oncology 500 v2 enable CGP from tissue samples. For other differences, see the comparison table on this page.
What analysis software can I use with TruSight Oncology 500 ctDNA v2 libraries?
The TruSight Oncology 500 ctDNA v2 analysis workflow uses an off-instrument software run on the DRAGEN v4 or v3 server or Illumina Connected Analytics platform to generate sample outputs, including high-level sample metrics, variants detected, and TMB and MSI scores. Tertiary analysis is enabled with either Illumina Connected Insights or the Velsera CGW.
Learn more about compatible analysis software products.
How long does TruSight Oncology 500 ctDNA v2 analysis take on Illumina Connected Analytics and a DRAGEN server?
Analysis on Illumina Connected Analytics takes about two hours for 4-8 libraries, three hours for 24 libraries, and approximately four hours for 64 libraries. Analysis on the DRAGEN server takes approximately three hours for 4 libraries, seven hours for 8 libraries, 19 hours for 24 libraries, and 43 hours for 64 samples.
Can I analyze TruSight Oncology 500 ctDNA v2 data with my own software?
Yes, you can perform analysis with your own software. However, Illumina will not be able to directly provide technical support in this case.
How much cfDNA input is required for TruSight Oncology 500 ctDNA v2?
Approximately 20 ng input cfDNA is recommended to achieve ~0.2% VAF detection for SNVs at >90% sensitivity and >95% specificity; however, the assay can accept a range of input from 10-30 ng cfDNA.
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