Pregled bibliografske jedinice broj: 65265
Detection of factor V Leiden by PCR-SSCP using precast Elchrom Scientific GMA gels
Detection of factor V Leiden by PCR-SSCP using precast Elchrom Scientific GMA gels // Biochemia Medica, vol 10, 6th Alps-Adria congress of clinical chemistry and laboratory medicine-abstracts / Suchanek, Ernest (ur.).
Zagreb: Hrvatsko društvo za medicinsku biokemiju i laboratorijsku medicinu (HDMBLM), 2000. (poster, domaća recenzija, sažetak, znanstveni)
CROSBI ID: 65265 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Detection of factor V Leiden by PCR-SSCP using precast Elchrom Scientific GMA gels
Autori
Ivanišević, Ana-Maria ; Štefanović, Mario ; Topić, Elizabeta
Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni
Izvornik
Biochemia Medica, vol 10, 6th Alps-Adria congress of clinical chemistry and laboratory medicine-abstracts
/ Suchanek, Ernest - Zagreb : Hrvatsko društvo za medicinsku biokemiju i laboratorijsku medicinu (HDMBLM), 2000
Skup
6th Alps-Adria congress of clinical chemistry and laboratory medicine
Mjesto i datum
Opatija, Hrvatska, 15.06.2000. - 17.06.2000
Vrsta sudjelovanja
Poster
Vrsta recenzije
Domaća recenzija
Ključne riječi
factor V Leiden; PCR-SSCP
Sažetak
Factor V Leiden point mutation (FV:R506Q) is the most common cause of thrombophilia. For the purpose of screening for this genetic abnormality in population at risk, a simple, reproducible and cheap method that would allow us to identify the carriers of Factor V mutation, is needed. The aim of this study was to optimize PCR-SSCP method for detection of Factor V Leiden mutation using the precast GMA gels in the Elchrom Scientific SEA 2000 apparatus. We performed PCR-SSCP analysis on 50 DNA samples previously genotyped by PCR-RFLP using Mnl I restriction enzyme. Exon X of Factor V was amplified using previously described primers and denatured at 95°C during 5 minutes. Denatured PCR products were run overnight. Gels were stained with SYBR Gold. We observed reproducible and uniform band patterns for mutant and wild type variants of Factor V. The concordance between PCR-SSCP and PCR-RFLP was 100 %. Technique has shown to be highly reproducible because of high performance characteristics of GMA gels and constant temperature maintenance by Elchrom Scientific SEA 2000 apparatus. In conclusion, described PCR-SSCP procedure is reliable, time saving and cost effective and should therefore be useful in the routine detection of Factor V Leiden.
Izvorni jezik
Engleski
Znanstvena područja
Temeljne medicinske znanosti
