An extended KRAS mutation test for the detection of 28 common mutations in FFPET and plasma specimens

Date

10 Oct 2016

Session

Poster display

Presenters

Jing Li

Citation

Annals of Oncology (2016) 27 (6): 15-42. 10.1093/annonc/mdw363

Authors

J. Li1, C. Hoeppner2, S. Gan1, A. Blair1, K. Min1, A. Sims2, A. Tietz2, M. Vinas2, T.T. Rehage2, K. Malhotra2, H. Halait2, V. Brophy1

Author affiliations

  • 1 Genomics And Oncology, Roche Molecular Systems, 94588 - Pleasanton/US
  • 2 Assay Development, Roche Molecular Systems, 94588 - Pleasanton/US
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Resources

Background

The KRAS oncogene is frequently mutated in human cancers. In addition to KRAS codons 12, 13, 61 hotspot mutations, recent clinical trial data revealed that mutations in KRAS codons 59, 117, 146 also predict poor response to anti-epidermal growth factor receptor (EGFR) therapy in patients with metastatic colorectal cancer (mCRC). Detection of an extended set of KRAS mutations in colorectal cancer tissues is now accepted clinical practice.

Methods

The KRAS Mutation Test v2 (Life Science Research, LSR) is a real-time PCR assay, which detects 28 mutations in codons 12, 13, 59, 61, 117 and 146 of the KRAS gene in DNA derived from formalin-fixed, paraffin-embedded tissue (FFPET) as well as plasma specimens. Mutations are detected by allele-specific amplification in three multiplex PCR reactions on the cobas z 480 analyzer. LOD studies were performed using contrived tissue and plasma samples. For method correlation, 301 FFPET specimens and 636 plasma specimens were tested with the KRAS Mutation Test v2 (LSR) and MiSeq (Illumina) sequencing as the reference method.

Results

Using preliminary analysis parameters, the KRAS test has shown it can detect at least 1% mutant sequence in a background of wild-type DNA for tissue, and at least 100 mutant sequence copies per mL for plasma in the LOD studies. Preliminary method correlation results revealed >98% overall concordance for tumor FFPET and >90% overall concordance for cell-free DNA samples with MiSeq reference data. Final results will be presented.

Conclusions

The KRAS Mutation Test v2 (LSR) demonstrated strong initial analytical performance for the detection of 28 KRAS mutations. It is a sensitive, robust and reliable assay with a quick turnaround time which can be used on both tissue and liquid biopsies.

Clinical trial identification

Legal entity responsible for the study

Roche Molecular Systems

Funding

Roche Molecular Systems

Disclosure

All authors have declared no conflicts of interest.

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