title: ÐÐµÑÐµÐºÑÐ¸Ñ Ð¼ÑÑÐ°ÑÐ¸Ð¹ Ð² Ð³ÐµÐ½Ð°Ñ KRAS, NRAS, BRAF Ð² Ð¾Ð±ÑÐ°Ð·ÑÐ°Ñ Ñ Ð¼Ð°Ð»ÑÐ¼ Ð¿ÑÐ¾ÑÐµÐ½ÑÐ½ÑÐ¼ ÑÐ¾Ð´ÐµÑÐ¶Ð°Ð½Ð¸ÐµÐ¼ Ð¾Ð¿ÑÑ Ð¾Ð»ÐµÐ²ÑÑ ÐºÐ»ÐµÑÐ¾Ðº Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ ÑÐ¸ÑÑÐ¾Ð²Ð¾Ð¹ ÐÐ¦Ð (dPCR)
title: Ð²ÑÐ¿ÑÑÐºÐ½Ð°Ñ ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð°Ñ ÑÐ°Ð±Ð¾ÑÐ° Ð±Ð°ÐºÐ°Ð»Ð°Ð²ÑÐ°
titleType: Subtitle
title: Detection of mutations in KRAS, NRAS, and BRAF genes in samples with a low percentage of tumor cells by digital PCR (dPCR)
Creators: name: ÐÑÐ·Ð¾Ð²ÐµÐ½ÐºÐ¾Ð²Ð°, ÐÐ°ÑÐ¸Ñ
Publisher: name: Ð¡Ð°Ð½ÐºÑ-ÐÐµÑÐµÑÐ±ÑÑÐ³ÑÐºÐ¸Ð¹ Ð¿Ð¾Ð»Ð¸ÑÐµÑÐ½Ð¸ÑÐµÑÐºÐ¸Ð¹ ÑÐ½Ð¸Ð²ÐµÑÑÐ¸ÑÐµÑ ÐÐµÑÑÐ° ÐÐµÐ»Ð¸ÐºÐ¾Ð³Ð¾
Year: 2022
Subjects: KRAS
NRAS
BRAF
Ð Ð¢Ð
Ð Ð
Ð¾Ð±ÑÐ°Ð·ÑÑ Ñ Ð¼Ð°Ð»ÑÐ¼ Ð¿ÑÐ¾ÑÐµÐ½ÑÐ½ÑÐ¼ ÑÐ¾Ð´ÐµÑÐ¶Ð°Ð½Ð¸ÐµÐ¼ Ð¾Ð¿ÑÑ Ð¾Ð»ÐµÐ²Ð¾Ð¹ ÑÐºÐ°Ð½Ð¸
ÑÐ¸ÑÑÐ¾Ð²Ð°Ñ ÐÐ¦Ð
ÑÐ°ÑÑÑÐµ Ð¼ÑÑÐ°ÑÐ¸Ð¸
TAQMAN Ð¿ÑÐ¾Ð±Ñ
LC
Ð¡Ð¡
samples with low percentage of tumour tissue
digital PCR
frequent mutations
TAQMAN probes

description: Ð¢ÐµÐ¼Ð° Ð²ÑÐ¿ÑÑÐºÐ½Ð¾Ð¸Ì ÐºÐ²Ð°Ð»Ð¸ÑÐ¸ÐºÐ°ÑÐ¸Ð¾Ð½Ð½Ð¾Ð¸Ì ÑÐ°Ð±Ð¾ÑÑ: Â«ÐÐµÑÐµÐºÑÐ¸Ñ Ð¼ÑÑÐ°ÑÐ¸Ð¸Ì Ð² Ð³ÐµÐ½Ð°Ñ KRAS, NRAS, BRAF Ð² Ð¾Ð±ÑÐ°Ð·ÑÐ°Ñ Ñ Ð¼Ð°Ð»ÑÐ¼ Ð¿ÑÐ¾ÑÐµÐ½ÑÐ½ÑÐ¼ ÑÐ¾Ð´ÐµÑÐ¶Ð°Ð½Ð¸ÐµÐ¼ Ð¾Ð¿ÑÑÐ¾Ð»ÐµÐ²ÑÑ ÐºÐ»ÐµÑÐ¾Ðº Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ ÑÐ¸ÑÑÐ¾Ð²Ð¾Ð¸Ì ÐÐ¦Ð  (dPCR)Â». Ð ÑÐ¾Ð²ÑÐµÐ¼ÐµÐ½Ð½Ð¾Ð¸Ì Ð´Ð¸Ð°Ð³Ð½Ð¾ÑÑÐ¸ÐºÐµ ÑÐ°ÐºÐ° Ð»ÐµÐ³ÐºÐ¸Ñ Ð¸ ÑÐ°ÐºÐ° ÑÐ¾Ð»ÑÑÐ¾Ð¸Ì ÐºÐ¸ÑÐºÐ¸ Ð²Ð°Ð¶Ð½ÑÐ¼ Ð¿ÑÐ¾Ð³Ð½Ð¾ÑÑÐ¸ÑÐµÑÐºÐ¸Ð¼ ÑÐ°ÐºÑÐ¾ÑÐ¾Ð¼ Ð»ÐµÑÐµÐ½Ð¸Ñ ÑÐ²Ð»ÑÐµÑÑÑ ÑÑÑÐ°Ð½Ð¾Ð²Ð»ÐµÐ½Ð¸Ðµ Ð¼ÑÑÐ°ÑÐ¸Ð¾Ð½Ð½Ð¾Ð³Ð¾ ÑÑÐ°ÑÑÑÐ° Ð² Ð³ÐµÐ½Ð°ÑÂ KRAS, NRASÂ Ð¸Â BRAF. ÐÑÑÐ²Ð»ÐµÐ½Ð¸Ðµ Ð¼ÑÑÐ°ÑÐ¸Ð¸Ì Ð² ÑÑÐ¸Ñ Ð³ÐµÐ½Ð°Ñ Ð¼Ð¾Ð¶ÐµÑ Ð¿Ð¾Ð¼Ð¾ÑÑ Ð²ÑÐ°ÑÐ°Ð¼ Ð² ÐºÐ¾ÑÑÐµÐºÑÐ¸ÑÐ¾Ð²Ð°Ð½Ð¸Ð¸ Ð»ÐµÑÐµÐ½Ð¸Ñ Ð¿Ð°ÑÐ¸ÐµÐ½ÑÐ° Ð¸ Ð½Ð°Ð·Ð½Ð°ÑÐµÐ½Ð¸Ð¸ ÑÐ°ÑÐ³ÐµÑÐ½Ð¾Ð¸Ì ÑÐµÑÐ°Ð¿Ð¸Ð¸. Ð Ð¾Ð±ÑÐ°Ð·ÑÐ°Ñ Ñ Ð½Ð¸Ð·ÐºÐ¸Ð¼ Ð¿ÑÐ¾ÑÐµÐ½ÑÐ½ÑÐ¼ ÑÐ¾Ð´ÐµÑÐ¶Ð°Ð½Ð¸ÐµÐ¼ Ð¾Ð¿ÑÑÐ¾Ð»ÐµÐ²Ð¾Ð¸Ì ÑÐºÐ°Ð½Ð¸ Ð½Ðµ Ð²ÑÐµÐ³Ð´Ð° ÑÐ´Ð°ÐµÌÑÑÑ Ð²ÑÑÐ²Ð¸ÑÑ ÑÐ¾ÑÐµÑÐ½ÑÑ Ð¼ÑÑÐ°ÑÐ¸Ð¾Ð½Ð½ÑÑ Ð·Ð°Ð¼ÐµÐ½Ñ Ð² ÐÐÐ ÐºÐ»Ð°ÑÑÐ¸ÑÐµÑÐºÐ¸Ð¼Ð¸ Ð¼ÐµÑÐ¾Ð´Ð°Ð¼Ð¸ (ÑÐµÐºÐ²ÐµÐ½Ð¸ÑÐ¾Ð²Ð°Ð½Ð¸Ðµ Ð¿Ð¾ Ð¡ÑÐ½Ð³ÐµÑÑ, Ð°Ð»Ð»ÐµÐ»Ñ-ÑÐ¿ÐµÑÐ¸ÑÐ¸ÑÐµÑÐºÐ°Ñ ÐÐ¦Ð ), Ð¿Ð¾ÑÑÐ¾Ð¼Ñ Ð² Ð´Ð°Ð½Ð½Ð¾Ð¸Ì ÑÐ°Ð±Ð¾ÑÐµ Ð¼Ñ Ð²Ð¾ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð»Ð¸ÑÑ Ð±Ð¾Ð»ÐµÐµ ÑÑÐ²ÑÑÐ²Ð¸ÑÐµÐ»ÑÐ½ÑÐ¼ Ð¼ÐµÑÐ¾Ð´Ð¾Ð¼ â Ð¦Ð¸ÑÑÐ¾Ð²Ð¾Ð¸Ì ÐÐ¦Ð  (Digital PCR, dPCR),Â ÐºÐ¾ÑÐ¾ÑÑÐ¸Ì Ð´Ð°ÐµÑ Ð²Ð¾Ð·Ð¼Ð¾Ð¶Ð½Ð¾ÑÑÑ Ð¿ÑÐ¾Ð²ÐµÑÐºÐ¸ Ð¾Ð±ÑÐ°Ð·ÑÐ¾Ð², ÑÐ¾Ð´ÐµÑÐ¶Ð°ÑÐ¸Ñ Ð²ÑÐµÐ³Ð¾ 1-2%Â Ð¾Ð¿ÑÑÐ¾Ð»ÐµÐ²Ð¾Ð¸Ì ÑÐºÐ°Ð½Ð¸. Ð Ð´Ð°Ð½Ð½Ð¾Ð¼ Ð¸ÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ð¸ Ð±ÑÐ»Ð° ÑÐ¾Ð·Ð´Ð°Ð½Ð° ÑÐµÑÑ-ÑÐ¸ÑÑÐµÐ¼Ð° Ð´Ð»Ñ Ð¿ÑÐ¾Ð²ÐµÑÐºÐ¸ Ð½Ð°Ð»Ð¸ÑÐ¸Ñ Ð¼ÑÑÐ°ÑÐ¸Ð¸Ì Ð² 37 ÑÐ°ÑÑÑÑ ÑÐ»ÑÑÐ°ÑÑ Ð² Ð³ÐµÐ½Ð°ÑÂ KRAS, NRASÂ Ð¸Â BRAF:Â Ð±ÑÐ»Ð¸ Ð¾Ð¿ÑÐ¸Ð¼Ð¸Ð·Ð¸ÑÐ¾Ð²Ð°Ð½Ñ ÑÑÐ»Ð¾Ð²Ð¸Ñ ÐµÐµ Ð¿ÑÐ¾Ð²ÐµÐ´ÐµÐ½Ð¸Ñ Ð¸ ÑÐ¾Ð·Ð´Ð°Ð½Ñ Ð´Ð¸Ð·Ð°Ð¸ÌÐ½Ñ Ð¿ÑÐ°Ð¸ÌÐ¼ÐµÑÐ¾Ð² Ð¸ Ð¿ÑÐ¾Ð±.Â ÐÑÐ»Ð¾ Ð¿ÑÐ¾Ð°Ð½Ð°Ð»Ð¸Ð·Ð¸ÑÐ¾Ð²Ð°Ð½Ð¾ 1919 Ð¾Ð±ÑÐ°Ð·ÑÐ¾Ð² Ñ Ð´Ð¸Ð°Ð³Ð½Ð¾Ð·Ð¾Ð¼ Ð Ð¢Ð Ð¸Â 1566Â Ð¾Ð±ÑÐ°Ð·ÑÐ¾Ð² Ñ Ð´Ð¸Ð°Ð³Ð½Ð¾Ð·Ð¾Ð¼ Ð Ð, Ð¸Ð· Ð½Ð¸Ñ Ð±ÑÐ»Ð¾ Ð¾ÑÐ¾Ð±ÑÐ°Ð½Ð¾ 66 Ð¾Ð±ÑÐ°Ð·ÑÐ¾Ð² Ð Ð Ð¸ 65 Ð¾Ð±ÑÐ°Ð·ÑÐ¾Ð² Ð Ð¢Ð Ð±ÐµÐ· ÑÐ°Ð½ÐµÐµ Ð²ÑÑÐ²Ð»ÐµÐ½Ð½ÑÑ Ð¼ÑÑÐ°ÑÐ¸Ð¸Ì Ð² Ð³ÐµÐ½Ð°ÑÂ KRAS, NRAS, BRAFÂ Ñ Ð½Ð¸Ð·ÐºÐ¸Ð¼ Ð¿ÑÐ¾ÑÐµÐ½ÑÐ½ÑÐ¼ ÑÐ¾Ð´ÐµÑÐ¶Ð°Ð½Ð¸ÐµÐ¼ Ð¾Ð¿ÑÑÐ¾Ð»ÐµÐ²Ð¾Ð¸Ì ÑÐºÐ°Ð½Ð¸Â (&lt;10%).Â ÐÐ¾ÑÐ»Ðµ Ð´Ð²ÑÑÑÑÑÐ¿ÐµÐ½ÑÐ°ÑÐ¾Ð¸Ì Ð¿ÐµÑÐµÐ¿ÑÐ¾Ð²ÐµÑÐºÐ¸ Ð½Ð° ÑÐÐ¦Ð  Ð¸ ÐºÐ°Ð¿ÐµÐ»ÑÐ½Ð¾Ð¸Ì ÑÐÐ¦Ð  Ð²Â 7Â Ð¾Ð±ÑÐ°Ð·ÑÐ°Ñ Ð±ÑÐ» Ð¿Ð¾Ð´ÑÐ²ÐµÑÐ¶Ð´ÐµÐ½ Ð¼ÑÑÐ°ÑÐ¸Ð¾Ð½Ð½ÑÐ¸Ì ÑÑÐ°ÑÑÑ, ÑÑÐ¾ ÑÐ²Ð¸Ð´ÐµÑÐµÐ»ÑÑÑÐ²ÑÐµÑ Ð¾ ÑÐ¾Ð¼, ÑÑÐ¾ Ð¼ÐµÑÐ¾Ð´Ñ, Ð¾ÑÐ½Ð¾Ð²Ð°Ð½Ð½ÑÐµ Ð½Ð° ÑÐ¸ÑÑÐ¾Ð²Ð¾Ð¸Ì ÐÐ¦Ð , Ð¿Ð¾Ð·Ð²Ð¾Ð»ÑÑÑ Ð²ÑÑÐ²Ð»ÑÑÑ Ð² Ð¾Ð±ÑÐ°Ð·ÑÐ°Ñ Ñ Ð½Ð¸Ð·ÐºÐ¸Ð¼ ÑÐ¾Ð´ÐµÑÐ¶Ð°Ð½Ð¸ÐµÐ¼ Ð¾Ð¿ÑÑÐ¾Ð»ÐµÐ²Ð¾Ð¸Ì ÑÐºÐ°Ð½Ð¸ Ð¼ÑÑÐ°ÑÐ¸Ð¸, Ð¿ÑÐ¾Ð¿ÑÑÐµÐ½Ð½ÑÐµ Ð¿ÑÐ¸ Ð¸ÑÐ¿Ð¾Ð»ÑÐ·Ð¾Ð²Ð°Ð½Ð¸Ð¸ ÑÑÐ°Ð½Ð´Ð°ÑÑÐ½ÑÑ Ð¿Ð¾Ð´ÑÐ¾Ð´Ð¾Ð².Â Â 
description: The subject of the graduate qualification work is Â«Detection of mutations in KRAS, NRAS, and BRAF genes in samples with a low percentage of tumor cells by digital PCR (dPCR)Â». In modern lung cancer and colorectal cancer diagnostics an important prognostic factor for treatment is the identification of mutational status in the KRAS, NRAS and BRAF genes. The detection of mutations in these genes can help doctors to adjust patient treatment and prescribe targeted therapy. It is not always possible to detect point mutation substitutions in DNA of samples with a low percentage of tumour tissue by classical methods (Sanger sequencing, allele-specific PCR), so in this study we used a more sensitive method - Digital PCR (dPCR), which allows us to test samples containing only 1-2% tumour tissue. In this study, a test system was established to check for the presence of mutations in 37 frequent cases in the KRAS, NRAS and BRAF genes: its conditions were optimised and primer and sample designs were created. A total of 1919 samples diagnosed with CC and 1566 samples diagnosed with LC were analysed, from which 66 samples of LC and 65 samples of CC without previously identified mutations in KRAS, NRAS, BRAF genes with low percentage of tumour tissue (&lt;10%) were selected. After two-step re-testing by PCR and droplet PCR, mutation status was confirmed in 7 samples, indicating that methods based on digital PCR allow detection of mutations missed by standard approaches in samples with low tumour tissue content.Â