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.

Objective Amenorrhea is a rare reproductive medical condition defined by the absence of menstruation during puberty or later life. This study aims to establish the frequency and pattern of chromosomal abnormalities (CA) in both primary amenorrhea (PA) and secondary amenorrhea (SA), and further to detect the genetic changes in exon 10 at nucleotide positions 919 and 2039 of the genotypes Thr307Ala, and Asn680Ser, respectively. Design, settings and patients This cross-sectional study was conducted on a sample of seventy amenorrhoeic women according to the Helsinki declaration rules of medical ethics, as divided into 40 (57.14%) with PA and 30 (42.86%) with SA, and 30 healthy women with normal menstruation as the control. The chromosomal karyotyping was performed according to the ISCN, 2020. PCR products were submitted to RFLP and Sanger sequencing for women with normal karyotype and high FSH serum levels. Results The classical Turner Syndrome was the most common CA in PA, followed by isochromosome X [46, Xi(X)(q10)], mosaicism of Turner and isochromosome X [45, X /46, Xi(X)(q10)], sex reversal (46, XY) and (46, XX,-3,+der3,-19,del 19 p). Abnormal SA cases were characterized by mosaicism Turner syndrome (45,X/46,XX) and (46,XX,-3,+der3,X,+derX). The homozygous genotypes AA and GG of Ala307Thr (rs6165) in the FSHR gene are most common in PA, while the homozygous genotype AA is more common in SA. GG and AG genotypes of Ser680Asn (rs6166) are more frequent in Iraqi patients with PA and SA compared to the healthy control women. Both PCR-RFLP and Sanger sequencing indicated a marked matching between genotypes. Conclusions The study emphasizes the need for cytogenetic analysis to determine the genetic basis of PA and SA. Further, genotyping for women with normal karyotype and high FSH serum concentrations via PCR-RFLP should be considered for the precise diagnosis and development of appropriate management of and counselling for these patients.