The right answers to frequently asked questions
Find the answers to all our products and services by clicking the links below.
General Questions regarding Sample Submission, Ordering & Sample Shipment & Results can be found here:
What kind of samples can be processed with INVIEW Oncoprofiling?
For the INVIEW Oncoprofiling, we accept fresh-frozen and FFPE tissue and genomic DNA.
For liquid biopsy analysis of cell-free DNA (cfDNA) isolated from blood plasma, please refer to our INVIEW Liquid Biopsy Oncoprofiling product.
How is the target enrichment performed?
The genes of interests are enriched via a proprietary Eurofins protocols. Target enrichment approach outcompetes common PCR-based enrichment with very uniform coverage and all exons of the genes covered. The proven target enrichment method has been optimised by Eurofins to develop highly complex libraries from low DNA and to deliver deep and uniform sequence coverage. Following target capture, next-generation sequencing of the library is performed.
What kind of mutations can be detected with INVIEW Oncoprofiling?
The service fully covers the entire exons of about 728 cancer-relevant genomic regions, including protein-coding genes, select promoter regions, miRNAs, and extra-exonic variants. In addition to SNPs and InDels, the product can also detect CNVs.
How is the tumor mutational burden (TMB) calculated?
- TMB is defined as the number of somatic, coding, base substitution, and InDel mutations per megabase of genome examined. All base substitutions and InDels in the coding region of targeted genes, including synonymous mutations, are initially counted before filtering as outlined below.
- The following mutations are excluded in silico from the TMB computation: Known somatic mutations in COSMIC and ClinVar, low-confident mutations, known germline variants in the ExAC (gnomAD) database, mutations predicted to be germline by the somatic-germline-zygosity algorithm, and mutations in tumor suppressor genes due to the focus of the Oncopanel All-In-One on actionable cancer mutations and potential panel design bias.
- To calculate the TMB per megabase, the total number of mutations counted is normalized by the size of the coding region of the targeted region in megabase (mutations per megabase, mut/MB). Due to the lack of standardization in TMB quantification we provide three TMB values:
- Non-synonymous mutations
- Non-synonymous mutations and plus indels and iii) including all mutations.
Is a focused gene panel, like INVIEW Oncoprofiling, suitable for TMB determination in comparison to whole-exome sequencing (WES)?
Although the gold standard for TMB determination is whole-exome sequencing (WES), focused panels, like the INVIEW Oncoprofiling, have been evaluated extensively for TMB estimation because of their lower sequencing cost and higher sensitivity. Recent research shows, that panels with a size > 1.5 Mbp are sufficiently suited to determine TMB with a high concordance to WES (Buchhalter et al. 2019, Int J Cancer 144:848–858). Thus, INVIEW Oncoprofiling's size of 3 Mbp provides more precise TMB estimates than other smaller panels.
What is the sensitivity of the product?
SNP and InDel detection has a technical sensitivity down to 1%.
What kind of quality controls do you perform?
The quality and quantity of each sample will be determined at sample receipt or after DNA extraction. Further quality controls are performed at various steps of the process.
What is the difference between INVIEW Oncoprofiling and INVIEW Liquid Biopsy Oncoprofiling?
Both tests offer cancer variant detection services based on a validated oncology panel. The interrogated genes and genomic alterations of the panel are the same. Similarly, the techniques used for both products are based on hybridisation for target enrichment and next-generation sequencing.
The two products differ in their starting material. INVIEW Oncoprofiling can accept conventional source material like fresh-frozen and FFPE tissues, whereas INVIEW Liquid Biopsy Oncoprofiling analyses cfDNA extracted from blood plasma samples.
Can I directly compare results between INVIEW Oncoprofiling and INVIEW Liquid Biopsy Oncoprofiling?
Yes, the two services are ideally suited for studies aimed at comparing the performance of traditional biopsies versus liquid biopsies.
What are the limitations of the CNV analysis?
Even when matched samples (tumour tissue and normal tissue or blood from the same patient) are submitted, the paired analysis approach could be subjected to limitations. The measurement variance on this single matched reference sample will be higher than that for a reference consisting of multiple reference samples. This could cause a modest number of (false-positive) CNV calls even in cases where the two paired samples are truly copy number identical.
Will all variants in my sample be detected?
Tumour heterogeneity could make it difficult to correctly identify all relevant copy number variations in a given sample. Even when the tumour itself is homogeneous, excessive contamination of normal cells within a tumour sample could lead to coverage differences that are too small to be detected even with Eurofins Genomics highly sensitive methods.
Can I use INVIEW Oncoprofiling for diagnostic purposes?
The product is available for research use only (RUO). The service is performed in an ISO17025:2005 –accredited and ISO13485:2016-certified laboratory.