Naslov
Simultaneous loss of heterozygosity and point mutation detection with high resolution melting

Autori
Musani, Vesna ; Sabol, Maja ; Ozretić, Petar ; Car, Diana ; Leović, Dinko ; Levacic Cvok, Mirela ; Levanat, Sonja

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Eur J Hum Genet / Van Ommen, G-J B - : Nature publishing group, 2011, 438-438

Skup
European Human Genetics Conference 2011

Mjesto i datum
Amsterdam, Nizozemska, 28.05.2011. - 31.05.2011

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
PTCH1; HRM; LOH

Sažetak
Basal-cell carcinoma (BCC) is the most common type of skin cancer, and shows a continuously increasing incidence. BCCs occur predominantly on head and neck of elderly fair-skinned people, with increased occurrence on the trunk (torso) in recent years. Patched (PTCH1) gene is a tumor suppressor gene most often associated with development of BCCs. It is a member of the Hh-Gli signaling pathway, a pathway that plays a major role in embryonic development and stem cell maintenance. Loss of heterozygosity (LOH) of tumor suppressor genes is a frequent event in tumorigenesis, and is most often analyzed by microsatellite typing. Here we offer a fast and efficient method for simultaneous SNP genotyping and mutation scanning, which can also be used for LOH detection. High resolution melting (HRM) analysis is based on differences in melting curves caused by variations in nucleotide sequence during the heating (melting) of the PCR product in the presence of the intercalating dye. Based on our previous experience with HRM, we expanded our mutation screening to LOH analysis with a study of 25 BCC samples. We also compared the LOH results obtained with HRM with those of the conventional microsatellite typing for several STR markers in PTCH1 region. HRM is an excellent method for both point mutation detection and LOH analysis, with point mutations found in 40%, and LOH in 50% of samples, and the interrater agreement between microsatellite and HRM LOH analyses of κ value of 0.869, what can be interpreted as very good agreement between the methods.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

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