ctDNA RAS Mutation Detection Kit

*Includes all controls.

Cell-free Mutation Detection

Mutations in KRAS and NRAS have been found in a number of malignancies, particularly metastatic colorectal cancer, lung adenocarcinoma, and thyroid cancer. Several studies have shown that specific mutations in KRAS and NRAS are less likely to respond to anti-EGFR therapies. [1, 2, 3]   Several diagnostics have been developed to identify such mutations. However, in a large number of cancer patients, tumor biopsy poses a significant health risk thereby limiting the tissue available for current molecular diagnostics. Non-invasive liquid biopsy diagnostics are revolutionizing cancer treatment selection, patient prognosis, and monitoring. Currently available qPCR kits for detecting RAS mutations have a limited sensitivity and coverage since they are not optimized for liquid biopsy purposes.

Recently, more studies have been focusing on the significance of KRAS mutation detected in cell-free DNA. Even though additional data are needed, a meta-analysis showed that KRAS mutation in cfDNA of cancer patient seems to act as a survival prognostic biomarker. [4]

The ctDNA RAS Mutation Detection Kit is a non-invasive, ultra-sensitive test that selectively amplifies and detects 25 clinically significant KRAS exon 2 and NRAS exons 2 and 3 mutations from circulating cell-free DNA derived from human plasma.

Testing Procedure

The ctDNA RAS Mutation Detection Kit is a single tube polymerase chain reaction (PCR)-based assay that use allele-specific primers in a multiplex reaction to identify the presence of clinically-actionable KRAS and NRAS mutations.  The assay works by amplifying mutant-specific sequences in samples that contain a mixture of mutant and wild-type DNA and rely on fluorescent probes for detection.

The testing procedure involves three (3) simple steps:

1. Isolation of DNA from plasma.
2. Amplification using the provided reagents in the kit.
3. Data analysis and interpretation.

Equipment and Materials

The ctDNA RAS Mutation Detection Kit requires a real-time PCR instrument capable of detecting VIC, FAM and CY5 fluorescent probes simultaneously.

All reagents required for PCR amplification/detection, as well as validated reaction controls are included. Reagents for DNA isolation are sold separately.

Intended Use:

USA: Available for research use only (RUO). Not for use in diagnostic procedures.

Europe: Available for research use only (RUO). Not for use in diagnostic procedures.

References:

1. Andreyev, H.J., et al. (2001) Kirsten ras mutations in patients with colorectal cancer: the ‘RASCAL II’ study. Br J Cancer 85, 692-696
2. Esteller, M., et al. (2001) K-ras and p16 aberrations confer poor prognosis in human colorectal cancer. J Clin Oncol 19, 299-304
3. Douillard, JY. et al. (2013) Panitumumab-FOLFOX4 Treatment and RAS Mutations in Colorectal Cancer. N Engl J Med 369, 1023-1034
4. Zhuang, R, Li S, Li Q, Guo X, Shen F, Sun H, et al. (2017) The prognostic value of KRAS mutation by cell-free DNA in cancer patients: A systematic review and meta-analysis. PLoS One 12(8): e0182562.