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

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.

Základní údaje

Originální název

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

Autoři

VANOVA, Barbora (703 Slovensko), Michal KALMAN (703 Slovensko), Karin JASEK (703 Slovensko), Ivana KASUBOVA (703 Slovensko), Tatiana BURJANIVOVA (703 Slovensko), Anna FARKASOVA (703 Slovensko), Peter KRUŽLIAK (703 Slovensko, domácí), Dietrich BUSSELBERG (634 Katar), Lukas PLANK (703 Slovensko) a Zora LASABOVA (703 Slovensko)

Vydání

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

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

30212 Surgery

Stát vydavatele

Itálie

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 2.644

Kód RIV

RIV/00216224:14110/19:00110194

Organizační jednotka

Lékařská fakulta

DOI

http://dx.doi.org/10.1007/s10238-019-00545-y

UT WoS

000464855400007

Klíčová slova anglicky

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

Štítky

14110121, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 16. 7. 2019 16:22, Soňa Böhmová

Anotace

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.

Citovat

VANOVA, Barbora, Michal KALMAN, Karin JASEK, Ivana KASUBOVA, Tatiana BURJANIVOVA, Anna FARKASOVA, Peter KRUŽLIAK, Dietrich BUSSELBERG, Lukas PLANK a Zora LASABOVA. Droplet digital PCR revealed high concordance between primary tumors and lymph node metastases in multiplex screening of KRAS mutations in colorectal cancer. Clinical and Experimental Medicine. Milan: SPRINGER-VERLAG ITALIA SRL, 2019, roč. 19, č. 2, s. 219-224. ISSN 1591-8890. Dostupné z: https://dx.doi.org/10.1007/s10238-019-00545-y.

@article{1545973,
   author = {Vanova, Barbora and Kalman, Michal and Jasek, Karin and Kasubova, Ivana and Burjanivova, Tatiana and Farkasova, Anna and Kružliak, Peter and Busselberg, Dietrich and Plank, Lukas and Lasabova, Zora},
   article_location = {Milan},
   article_number = {2},
   doi = {http://dx.doi.org/10.1007/s10238-019-00545-y},
   keywords = {Colorectal cancer; KRAS mutation testing; Droplet digital PCR; Lymph nodes},
   language = {eng},
   issn = {1591-8890},
   journal = {Clinical and Experimental Medicine},
   title = {Droplet digital PCR revealed high concordance between primary tumors and lymph node metastases in multiplex screening of KRAS mutations in colorectal cancer},
   url = {http://dx.doi.org/10.1007/s10238-019-00545-y},
   volume = {19},
   year = {2019}
}

TY  - JOUR
ID  - 1545973
AU  - Vanova, Barbora - Kalman, Michal - Jasek, Karin - Kasubova, Ivana - Burjanivova, Tatiana - Farkasova, Anna - Kružliak, Peter - Busselberg, Dietrich - Plank, Lukas - Lasabova, Zora
PY  - 2019
TI  - Droplet digital PCR revealed high concordance between primary tumors and lymph node metastases in multiplex screening of KRAS mutations in colorectal cancer
JF  - Clinical and Experimental Medicine
VL  - 19
IS  - 2
SP  - 219-224
EP  - 219-224
PB  - SPRINGER-VERLAG ITALIA SRL
SN  - 15918890
KW  - Colorectal cancer
KW  - KRAS mutation testing
KW  - Droplet digital PCR
KW  - Lymph nodes
UR  - http://dx.doi.org/10.1007/s10238-019-00545-y
L2  - http://dx.doi.org/10.1007/s10238-019-00545-y
N2  - 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.
ER  -

VANOVA, Barbora, Michal KALMAN, Karin JASEK, Ivana KASUBOVA, Tatiana BURJANIVOVA, Anna FARKASOVA, Peter KRUŽLIAK, Dietrich BUSSELBERG, Lukas PLANK a Zora LASABOVA. Droplet digital PCR revealed high concordance between primary tumors and lymph node metastases in multiplex screening of KRAS mutations in colorectal cancer. \textit{Clinical and Experimental Medicine}. Milan: SPRINGER-VERLAG ITALIA SRL, 2019, roč.~19, č.~2, s.~219-224. ISSN~1591-8890. Dostupné z: https://dx.doi.org/10.1007/s10238-019-00545-y.

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