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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

Ivanišević, Ana-Maria ; Štefanović, Mario ; Topić, Elizabeta 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. str. 118-x

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

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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

Povezanost rada

Temeljne medicinske znanosti