Mutation analysis

Two main sequencing methods are used: Sanger sequencing and next-generation sequencing (NGS).

Material
Histological slides/blocks, cytological preparations and fresh tissue can be used for both methods.

Next-generation sequencing (NGS)

Nowadays, it is possible to detect all relevant mutations within a short time through the parallel sequencing of several genes on the tumour DNA and RNA, using high-throughput sequencing (so-called next-generation sequencing, or NGS). In particular, NGS makes it possible to conduct targeted DNA- and RNA-based sequencing for the simultaneous detection of clinically relevant mutations, copy number variations and fusion transcripts. It can also be used for small biopsies with low tumour cell content. This method produces robust results in routine diagnostics and meets modern requirements for treatment stratification.

The following panels are available in our range:

• Oncomine™ Comprehensive Assay Plus, incl. TMB, MSI, DNA (392 genes) / RNA (51 fusion genes)
• Oncomine™ Comprehensive v3 Assay, DNA (135 genes) / RNA (51 fusion genes)
• Oncomine™ Focus Assay DNA (35 genes) / RNA (23 fusion genes)
• Oncomine™ BRCA Panel (all coding sections of the BRCA1 and BRCA2 genes)
• Oncomine™ Tumor Mutation Load Assay (TMB, tumour mutation burden per megabase, 409 genes)
• Liquid Biopsy, Oncomine™ Pan-Cancer Cell-Free Assay

Sanger sequencing

Sanger sequencing is one of the classic methods of DNA sequencing that makes it possible to determine the base order of short sequences of a specific gene. In tumour diagnostics, DNA sequences of sick patients are compared with DNA sequences of healthy individuals to identify mutated DNA sequences in the tumour.

We analyse the following genes using Sanger sequencing:

• BRAF (exon 15)
• EGFR (exon 18, 19, 20, 21)
• ERBB2/HER2 (exon 19, 20)
• KRAS (exon 2, 3, 4)
• KIT (exon 8, 9, 11, 13, 14, 17)
• NRAS (exon 2, 3, 4)
• PDGFRA (exon 12, 18)
• TERT promoter