Detection of factor V Leiden by PCR-SSCP using precast Elchrom Scientific GMA gels (CROSBI ID 479893)
Prilog sa skupa u zborniku | sažetak izlaganja sa skupa | domaća recenzija
Podaci o odgovornosti
Ivanišević, Ana-Maria ; Štefanović, Mario ; Topić, Elizabeta
engleski
Detection of factor V Leiden by PCR-SSCP using precast Elchrom Scientific GMA gels
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.
factor V Leiden; PCR-SSCP
nije evidentirano
nije evidentirano
nije evidentirano
nije evidentirano
nije evidentirano
nije evidentirano
Podaci o prilogu
118-x.
2000.
objavljeno
Podaci o matičnoj publikaciji
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)
Podaci o skupu
6th Alps-Adria congress of Clinical Chemistry and Laboratory Medicine
poster
15.06.2000-17.06.2000
Opatija, Hrvatska