VAL34LEU POLYMORPHISM DETECTION BY REAL TIME PCR ASSAY USING FLUORESCENCE RESONANCE ENERGY TRANSFER ON ROTOR-GENE 6000

Year 2016, Volume: 1 Issue: 1, 15 - 19, 26.01.2016

Maryam Naser Kamjoo , Ali Nazemı

https://doi.org/10.23884/ijhsrp.2016.1.1.02

Abstract

Factor XIII (FXIII) Val34Leu is the most important

polymorphism of the A subunit in factor XIII gene. The aim of the present

study was to develop an efficient method based on real time PCR with

Fluorescence resonance energy transfer (FRET) detection and melting curve

analysis for the detection of Val34Leu polymorphism.

Specific primers were used to amplify the relevant fragment of the factor

XIII-A gene and fluorescence resonance energy transfer hybridization

probes were used for detection in a Rotor-Gene Q 5Plex HRM platform.

Melting temperature (Tm) for the wild type was at 68.8, while Tm for

homozygote mutant was at 63.8; heterozygote demonstrated both peaks.

Our results showed that primers and probes propoesd in this study

demonstrated a high specificity to identify wild type , heterozygous and

homozygous mutantgenotypes.

Due to the increasing molecular diagnosis in developing countries and the

importance of identifying polymorphisms, this real time PCR assay is of

great importance. An important advantage of this approach is the high

sensitivity and specificity.

Keywords

factor XIII, val34leu, Fluorescence resonance energy transfer

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