TruSight Tumor 170

Comprehensive next-generation sequencing (NGS) assay that targets 170 genes associated with solid tumors.

~2 days

Assay time

~10.5 hr

Hands-on time

40 ng DNA and/or RNA

Input quantity

See full details in the specifications table

The TruSight Tumor 170 bundle kits that contain NextSeq 500/550 v2 sequencing reagents (Catalog nos. OP-101-1003 and 20018621) have been discontinued. The TruSight Tumor 170 Kit with NextSeq v2.5 Reagents (Catalog no. 20028821) is the recommended replacement product. The TruSight Tumor 170 Kit Plus Watson for Genomics (Catalog no. 20018622) has also been discontinued. If you require that analysis, continue to work with IBM directly. Illumina remains committed to providing you with high-quality support and service.

Overview

TruSight Tumor 170 is an NGS assay that assesses 170 genes associated with common solid tumors.

  • Assess fusions, splice variants, insertions/deletions (indels) and single nucleotide variants (SNVs), and amplifications in one assay using DNA and RNA

  • Detect variants at 5% allele frequency with as little as 40 ng DNA and RNA input

  • Maximize results with reliable small variant detection from high- and low-quality FFPE samples

Comprehensive coverage of cancer-related variants

Maximize single-assay efficiency using DNA and RNA from a single sample to assess multiple variant types, preserving valuable tissue, time, and resources. Expert-selected evidence-based content provides researchers with comprehensive coverage of variants most likely to play a role in tumorigenesis.

Combined workflow for DNA and RNA

DNA and RNA libraries are prepared, sequenced, and analyzed simultaneously for efficient assessment of numerous types of somatic variants.

Streamlined data analysis

Use the TruSight Tumor 170 App on BaseSpace Sequence Hub for sequencing data analysis and management. Local secondary analysis is available on a Docker-based software. 


Specifications


Required products

Requires use of NextSeq 500/550 v2.5 reagents and Pierian interpretation software.

/ Results

Applications

TruSight Tumor 170 provides efficient comprehensive coverage of cancer-related variants (small variants, amplifications, splice variants, and fusions) in RNA and DNA obtained from a single FFPE sample.

Example workflow



Project recommendations

Instrument Recommended number of samples Read length
NextSeq 550 System

Samples per run (high output): 16 (8 DNA + 8 RNA), 10 if DNA only, 16 if RNA only

2 × 101 bp (max recommended)


Related applications and methods

Compare

TruSight Tumor 170 TruSight Oncology 500 High-Throughput TruSight Oncology 500 TruSight Oncology 500 ctDNA v2
Assay time ~2 days

4–5 days from sample to results

4–5 days from sample to results

3-4 days from purified nucleic acid to variant report

Automation details Explore available automation methods Explore available automation methods Explore available automation methods
Cancer type Solid Tumor Pan-Cancer, Solid Tumor Pan-Cancer, Solid Tumor Pan-Cancer, Solid Tumor
Content specifications Probes enrich for full coding sequences of 170 genes. Calls single nucleotide variants, small insertions, and deletions in 151 genes, amplifications in 59 genes, and fusions plus splice variants in 55 genes.

Targeted sequencing of DNA from 523 genes of interest and RNA from 55 genes, for a total of 1.94Mb panel size. MSI and TMB measurement included. 

Targeted sequencing of DNA from 523 genes and RNA from 55 genes for a total of 1.94 Mb panel size. MSI and TMB measurement included. 

The optional TruSight Oncology 500 HRD kit (not available in Japan) content includes coverage of ~25K SNPs to assess homologous recombination deficiency through a comprehensive genomic instability score (LOH+TAI+LST) powered by Myriad Genetics. 

Targeted selection of 523 genes (full coding sequence) for a total of 1.94Mb panel size.
• Immuno-oncology biomarker coverage: TMB and MSI
• Guideline coverage: Broad coverage of key guidelines for multiple solid tumor types
• Clinical trial coverage: Over 600 clinical trials (based on Pierian clinical knowledgebase, as of February 2023)
Description Perform comprehensive somatic variant detection research in solid tumors using variant calling information from both DNA and RNA.

A high-throughput comprehensive NGS assay to identify key biomarkers in guidelines and > 1K clinical trials from a streamlined workflow using the NovaSeq 6000 System or NovaSeq 6000Dx Instrument (in Research Mode). Includes coverage of immuno-oncology biomarkers MSI and TMB.

Assay that enables comprehensive genomic profiling from FFPE tissue and runs on the NextSeq 550 System or NextSeq 550Dx Instrument (in Research Mode) and can batch up to eight samples at a time.

Provides a noninvasive method for profiling solid tumors for cancer research applications through comprehensive genomic profiling of liquid biopsy samples (ctDNA from blood plasma). This liquid biopsy approach provides insights about intra- and inter-tumor heterogeneity using a minimally invasive sample collection approach to complement tissue-based CGP.

Hands-on time ~10.5 hr ~2.5 hr for automated workflow
~10.5 hr for manual workflow
~2.5 hr for automated workflow
~10.5 hr for manual workflow

~1.5-2.5 hr for library prep and enrichment

Input quantity 40 ng DNA and/or RNA

40 ng DNA and/or 40–80 ng RNA

40 ng DNA and/or 40 ng RNA

20 ng cfDNA (4 ml of plasma)

Instruments NextSeq 550 System, NextSeq 500 System, HiSeq 2500 System NovaSeq 6000Dx in Research Mode, NovaSeq 6000 System NextSeq 550 System, NextSeq 550Dx in Research Mode, NextSeq 500 System NovaSeq 6000 System
Method Targeted DNA sequencing, Targeted RNA sequencing, Target enrichment Targeted DNA sequencing, Targeted RNA sequencing, Target enrichment Targeted DNA sequencing, Targeted RNA sequencing, Target enrichment Targeted DNA sequencing, Target enrichment
Multiplexing Up to 32-plex for DNA using both index sets, up to 16-plex for RNA

Up to 16-plex (SP flow cell), 32-plex (S1 flow cell), 72-plex (S2 flow cell), and 192-plex (S4 flow cell)

Up to 8-plex Up to 8 on S2, and 24 on S4 using Xp-4 Lane workflow, 16 indexes maximum
Nucleic acid type DNA, RNA DNA, RNA DNA, RNA DNA
Specialized sample types Low-Input Samples, FFPE Tissue FFPE Tissue FFPE Tissue Circulating Tumor DNA, Blood
Species category Human Human Human Human
Technology Sequencing Sequencing Sequencing Sequencing
Variant class Gene fusions, Somatic variants, Structural variants, Single nucleotide variants (SNVs), Insertions-deletions (indels), Copy number variants (CNVs) Gene fusions, Somatic variants, Novel transcripts, Structural variants, Transcript variants, Single nucleotide variants (SNVs), Insertions-deletions (indels), Copy number variants (CNVs) Gene fusions, Transcript variants, Single nucleotide variants (SNVs), Insertions-deletions (indels), Copy number variants (CNVs), Loss of heterozygosity (LOH), Somatic variants, Novel transcripts, Single nucleotide polymorphisms (SNPs), Structural variants Single nucleotide variants (SNVs), Insertions-deletions (indels), Copy number variants (CNVs)

Selection tools:

Figures

Gene content in the TruSight Tumor 170 assay

TruSight tumor 170 targets all coding exons per the current RefSeq database1 in 170 genes. The content includes 55 genes for fusions and splice variants, 148 SNVs and indels, and 59 amplifications.

Combined library prep workflow

DNA and RNA samples follow the same workflow, after the cDNA synthesis step (for RNA) and the shearing step (for DNA).

Target coverage from FFPE samples

DNA from FFPE samples of varying quality was extracted and evaluated using TruSight Tumor 170 and sequenced on the NextSeq 500 System. Sample quality was assessed using qPCR to measure DNA amplification potential. The ΔCq value indicates the cycle threshold (Ct) value of each DNA sample minus the Ct value of a DNA standard.

Resources

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Learn how a single assay interrogates both DNA and RNA to maximize the output from a single FFPE sample.

TruSight Tumor 170 Kit (24 Samples)

OP-101-1004

Includes library prep and enrichment reagents to allow for solid tumor profiling of 170 genes from FFPE samples.

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TruSight Tumor 170 Kit, With NextSeq v2.5 Reagents (24 Samples)

20028821

Includes library prep, enrichment, and sequencing consumables to allow for solid tumor profiling of 170 genes from FFPE samples.

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FAQs

During library prep and enrichment, the DNA and RNA samples go through the same workflow following sample shearing (DNA) and cDNA synthesis (RNA). This results in the ability to process the DNA and RNA samples in parallel. The DNA and RNA libraries can then be sequenced on the same sequencing run.

Extraction reagents are not included in the TruSight Tumor 170 kit. The QIAGEN AllPrep DNA/RNA FFPE Kit (Catalog no. 80234) has demonstrated high yields of nucleic acids in comparison to other extraction methods for this assay. The QIAGEN AllPrep DNA/RNA FFPE Kit includes a DNase I digestion step during RNA extraction. You can select other commercially available extraction kits at your discretion.

TruSight Tumor 170 performance has been verified on the NextSeq 500 and NextSeq 550 Systems, and on the HiSeq 2500 System in rapid run mode.

Illumina recommends adding a minimum of 40 ng of DNA or RNA. For samples that are highly degraded, assay performance may be improved by using a maximum of 120 ng of DNA and 85 ng of RNA.

Coverage may vary based on sample quality and multiplexing level. Please review the DNA_SampleMetricsReport.txt and RNA_SampleMetricsReport.txt files produced by the TruSight Tumor 170 app for each analysis. This report contains a metric indicating the percent bases with > 100× coverage. A sequencing run with 8 high-quality DNA and 8 high-quality RNA samples (16 libraries) has demonstrated ≥ 99% bases with 100× coverage.

Use the TruSight Tumor 170 app available in BaseSpace Sequence Hub cloud environment or a local Docker App to analyze TruSight Tumor 170 libraries.

/ Results

Order the NovaSeq X Series

Advanced chemistry, optics, and informatics combine to deliver exceptional speed and data quality, outstanding throughput and scalability.

MiSeq Reagent Kit v3

Optimized reagent kit that provides increased cluster density and read length, improving sequencing quality scores compared to earlier versions.

TruSight Oncology 500

Enable CGP with a large pan-cancer panel covering all major variant classes plus gene signatures (TMB, MSI, and HRD) from FFPE tissue.

Illumina DNA Prep

A fast, integrated workflow for preparing libraries for use in a wide range of sequencing applications.

References

1. O'Leary NA, Wright MW, Brister JR, et al. Reference sequence (RefSeq) database at NCBI: current status, taxonomic expansion, and functional annotation. Nucleic Acids Res. 2016;44(D1):D733-45.

 doi:10.1093/nar/gkv1189

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