STUDY ON IDENTIFYING THE C677T POLYMORPHISM OF MTHFR GENE BY PCR-RFLP TECHNIQUE IN PATIENTS WITH ABNORMAL SEMEN ANALYSIS

Abstract

Background: The C677T polymorphism of MTHFR gene is a risk factor of many diseases, including male infertility. This study is aimed at: 1. Improving a PCR-RFLP process with the own designed primers to identify the C677T polymorphism of MTHFR gene. 2. Evaluating the prevalence of the C677T polymorphism of MTHFR gene in the volunteer group and the male with abnormal semen analysis. Materials and Method: DNA samples was extracted from peripheral blood of 60 volunteers and 30 patients with abnormal semen analysis. Designing primers by using FastPCR software, then improving PCR-RFLP technique. Confirming the results of polymorphism by DNA sequencing technique. Evaluating the prevalence of the C677T polymorphism of MTHFR gene. Results: We designed successfully primers to amplify fragment of MTHFR gene including C677T polymorphism and an obligatory restriction site of HinfI (as internal control). The MTHFR genotype frequencies in the volunteer group were: 71.67% (677CC); 25% (677CT); and 3.33% (677TT). Genotype frequencies in patients with abnormal semen analysis were 66.67% ; 30% and 3.33% ; in group of azoospermia were 53.34% ; 40% and 6.66% ; in group of oligospermia were 80%; 20% and 0%. Conclusion: We standardized successfully PCR-RFLP technique to identifying C677T polymorphism of MTHFR gene.We evaluated the prevalence of the C677T polymorphism of MTHFR gene in the volunteer group and the male with abnormal semen analysis.