J 2019

Droplet digital PCR revealed high concordance between primary tumors and lymph node metastases in multiplex screening of KRAS mutations in colorectal cancer

VANOVA, Barbora, Michal KALMAN, Karin JASEK, Ivana KASUBOVA, Tatiana BURJANIVOVA et. al.

Basic information

Original name

Droplet digital PCR revealed high concordance between primary tumors and lymph node metastases in multiplex screening of KRAS mutations in colorectal cancer

Authors

VANOVA, Barbora (703 Slovakia), Michal KALMAN (703 Slovakia), Karin JASEK (703 Slovakia), Ivana KASUBOVA (703 Slovakia), Tatiana BURJANIVOVA (703 Slovakia), Anna FARKASOVA (703 Slovakia), Peter KRUŽLIAK (703 Slovakia, belonging to the institution), Dietrich BUSSELBERG (634 Qatar), Lukas PLANK (703 Slovakia) and Zora LASABOVA (703 Slovakia)

Edition

Clinical and Experimental Medicine, Milan, SPRINGER-VERLAG ITALIA SRL, 2019, 1591-8890

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

30212 Surgery

Country of publisher

Italy

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

Impact factor

Impact factor: 2.644

RIV identification code

RIV/00216224:14110/19:00110194

Organization unit

Faculty of Medicine

DOI

UT WoS

000464855400007

Keywords in English

Colorectal cancer; KRAS mutation testing; Droplet digital PCR; Lymph nodes

Tags

Tags

International impact, Reviewed
Změněno: 16/7/2019 16:22, Soňa Böhmová

Abstract

V originále

The proto-oncogene KRAS belongs among the most frequently mutated genes in all types of cancer and is also very important oncogene related to colorectal tumors. The detection of mutations in this gene in primary tumor is a predictive biomarker for the anti-EGFR therapy in metastatic CRC (mCRC); however, the patients with wild-type KRAS can also show resistance to the personalized medicine. The droplet-based digital PCR technology has improved the analytical sensitivity of the mutations detection, which led us to the idea about the optimization of this approach for KRAS testing. In this study, we report the application of ddPCR technology in order to analyze the presence of KRAS mutations in primary tumor and matched metastasis in lymph nodes (LNs) from patients with mCRC and address the question, whether the improvement in the detection method can lower the discrepancies of KRAS mutations detection between the primary tumor and regional LNs. Genomic DNA with wtKRAS and commercial DNA with mtKRAS (G12D) were used to set up the ddPCR reaction. Formalin-fixed paraffin-embedded tissues from primary tumor and positive lymph node from 31 patients with mCRC were analyzed using ddPCR and Sanger sequencing. KRAS status of primary tumors was known; however, the mutation status of lymph nodes was not detected previously. From 31 samples of primary tumors, our results corresponded to results from IVD kit in 30 cases. For one patient, ddPCR detected KRAS mutation in comparison with negative result of the IVD kit. In the samples of metastatic infiltrated LNs, ddPCR detected 16 samples as a WT KRAS and 15 lymph nodes showed positivity for KRAS mutation, whereby Sanger sequencing found KRAS mutations in 8 cases only. We also found two cases where genetic conditions of KRAS gene differed between primary tumor and infiltrated lymph node, both "low-grade" adenocarcinoma. Our study approved that ddPCR method is adequate technique with high sensitivity and in the future may be used as a diagnostic tool for evaluation of KRAS mutations, especially in infiltrated LNs of patients with mCRC.