In terms of the PCR-RFLP technique, genetic screening of Ala575Val inactivating mutation in patients with amenorrhea

Abstract

When mutation is inactivated in the follicle-stimulating hormone receptor (FSHR) gene of patients with amenorrhea, the receptor’s functionality is abolished by completely blocking it, altering the ligand–receptor complex or altering the essential hormone signal transduction. This study aims to detect the frequency and pattern of chromosomal abnormalities and the presence of inactivating mutations (Ala575Val) at position 1540 of the FSHR gene in Iraqi women diagnosed with secondary amenorrhea (SA) and primary amenorrhea (PA). This cross-sectional study was carried out between February and August 2022. Depending on the initial diagnosis of amenorrhea, women were split into two groups: PA and SA. The polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) was done for those women with normal karyotyping and hypergonadotropic hypogonadism [follicle stimulating hormone (FSH) > 20 mIU/ml] to determine the genetic cause, such as inactivating mutation in Exon 10E. The FSH of blood in study patients with PA was 72.28 ± 23.60, luteinizing hormone (LH) was 20.60 ± 13.55, and BMI was 24.012 ± 4.06. In SA, the FSH, LH, and BMI values were 69.821 ± 35.95, 16.788 ± 14.12, and 26.12 ± 4.37, respectively. Also, highly significant differences were observed compared with healthy control subjects (P-value < 0.005). The high matching between the results obtained by PCR-RFLP and those obtained by Sanger sequencing techniques used in this study confirmed no detected mutation in Ala575Val at position 1540 in all patients with PA and SA. The study concluded that in the PCR study, Ala575Val encoding genes are highly detected, while in PCR-RFLP, no action of MscI restriction enzyme in position 1540 (region of Ala575Val genotype) has emerged. This gives the impression that women with amenorrhea in the Iraqi population might not have any inactivating mutations in the FSHR gene.