COMPARISON OF DIFFERENT METHODS OF MOLECULAR-GENETIC ANALYSIS OF SOMATIC MUTATIONS IN K-RAS GENE IN PATIENTS WITH COLORECTAL CANCER

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Abstract

Two approaches to somatic point mutations in 12 and 13 codones of K-ras gene were analyzed: PCR/SSCP/AСRS/sequencing and allele-specific PCR in the real-life regimen (Russian set «KRAS-7M»). The comparison was carried out on 62 examples of genomic DNA extracted from frozen colon carcinomas, which underwent manual dissection. The results obtained in two attempts were consistent in 95,2% (N=59). Specificity and sensitivity of K-ras mutations detection using «KRAS-7M» set were 100 and 96,4% respectively, and 94,1 and 100% respectievly using PCR/SSCP/AСRS/ automatic sequencing. False positive results were absent when detecting with «KRAS-7M» and accounted for 2 cases (5,9%) when using PCR/SSCP/ AСRS/automatic sequencing. The only false negative response (3,6%) was obtained analyzing mutations using «KRAS-7M».

About the authors

F. A. Amosenko

Scientific centre of medical genetics, RAMS, Moscow
Blokhin Russian oncological scientific centre RAMS, Moscow

Author for correspondence.
Email: amossenko@med-gen.ru
кандидат биологических наук, старший научный сотрудник, ведущий научный сотрудник Медико-генетического научного центра РАМН, лаборатории ДНК-диагностики Адрес: 115478, Москва, ул. Москворечье, д. 1 Тел.: (965) 132-78-02, факс: (499) 324-07- 02 Россия

I. V. Karpov

Laboratory of genetic diagnosis «BioLink», Novosibirsk

Email: biolink@inbox.ru
лаборант ООО «БиоЛинк» лаборатории генодиагностики Адрес: 630117, Новосибирск, ул. Академика Тимакова, д. 2 Тел./факс: (383) 334-86-14 Россия

A. V. Polyakov

Scientific centre of medical genetics, RAMS, Moscow

Email: info@dnalab.ru
доктор биологических наук, профессор Медико-генетического научного центра РАМН, заведующий лабораторией ДНК-диагностики Адрес: 115478, Москва, ул. Москворечье, д. 1 Тел./факс: (495) 221-90-84 Россия

S. P. Kovalenko

Institute of molecular biology and biophysics Siberian division of RAMS, Novosibirsk

Email: sp_kovalenko@yahoo.com
кандидат биологических наук, заведующий лабораторией генной инженерии Института молекулярной биологии и биофизики Сибирского Отделения РАМН Адрес: 630117, Новосибирск, ул. Академика Тимакова, д. 2 Тел/факс: (383) 333-47-10 Россия

V. A. Shamanin

Laboratory of genetic diagnosis «BioLink», Novosibirsk

Email: va_shamanin@biolinklab.ru
кандидат биологических наук, ведущий научный сотрудник лаборатории гено- диагностики ООО «БиоЛинк» Адрес: 630117, Новосибирск, ул. Академика Тимакова, д. 2 Тел/факс: (383) 334-86-14 Россия

L. N. Lyubchenko

Blokhin Russian oncological scientific centre RAMS, Moscow

Email: clingen@mail.ru
доктор медицинских наук, заведующая лабораторией клинической онкогенетики Российского онкологического научного центра им. Н.Н. Блохина РАМН Адрес: 115478, Москва, Каширское шоссе, д. 24 Тел.: (916) 623-23-66, факс: (499) 324-96-29 Россия

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