Abstract
BackgroundPolycystic ovary syndrome (PCOS), the commonest endocrine disorder affecting young women, appears to be a multigenic trait with contributing genes being unclear. Hence, analysis of polymorphisms in multiple candidate genes is required. Currently available genotyping methods are expensive, time-consuming with limited analytical sensitivity.Aim(i) Develop and validate high resolution melting (HRM) assay and allele-specific real-time quantitative PCR (AS-qPCR) for genotyping selected SNPs associated with PCOS.(ii) Identify selected SNPs and their association with a Sri Lankan cohort of well-characterized PCOS.MethodsDNA was extracted from women with well-characterized PCOS from adolescence (n = 55) and ethnically matched controls (n = 110). FTO (Fat mass and obesity associated gene; rs9939609), FSHB (Follicle stimulating hormone beta subunit; rs6169), FSHR (Follicle stimulating hormone receptor; rs6165/rs6166), and INSR (Insulin receptor; rs1799817) genes were genotyped using HRM assay. GnRH1 (Gonadotropin releasing hormone; rs6185), LHB (Luteinizing hormone beta subunit; rs1800447/rs34349826) and LHCGR (Luteinizing hormone/choriogonadotropin receptor; rs2293275) genes were genotyped using AS-qPCR method. Genotyping results were validated using Sanger sequencing.ResultsA significant association was observed within FTO gene polymorphism (rs9939609) and PCOS. Genotype frequency of FTO gene (rs9939609)—cases versus controls were TT-36.4% vs.65.4% (p<0.05), AT-23.6% vs.20.9%, AA-40% vs.13.6% (p<0.05). Genotype frequencies of the SNPs GnRH1 (rs6185), FSHB (rs6169), FSHR (rs6165 & rs6166), LHB (rs1800447 & rs34349826), LHCGR (rs2293275) and INSR (rs1799817) were not significantly different between cases and controls (p>0.05). Only the mutant alleles were observed for LHB rs1800447 and rs34349826 SNPs in both groups. The HRM and AS-qPCR assay results had 100% concordance with sequencing results.ConclusionsFTO gene rs9939609 polymorphism is significantly more prevalent among Sri Lankan PCOS subjects while the other selected SNPs of HPG axis genes and INSR gene showed no association. HRM and AS-qPCR assays provide a reliable, fast and user-friendly genotyping method facilitating wider implication in clinical practice.
Highlights
Polycystic ovary syndrome (PCOS) is the commonest endocrine disorder affecting women of reproductive age with a prevalence varying between 5–13%
A significant association was observed within FTO gene polymorphism and PCOS
Brower et al evaluated whether the variants associated with PCOS in Han Chinese are associated with PCOS in white Europeans and concluded that DENND1A, THADA, FSHR, illustrates the amplification plot of SNP–rs2293275 (INSR) and YAP1 loci are likely to play important roles in the etiology of PCOS across populations [11]
Summary
Polycystic ovary syndrome (PCOS) is the commonest endocrine disorder affecting women of reproductive age with a prevalence varying between 5–13%. Some recent studies have identified varying SNP sites to be associated with PCOS in differing populations. The recent Genomewide association study (GWAS) by Chen et al showed 3 SNP sites on THADA, DENND1A and TOX3 to be associated with PCOS [10]. Brower et al evaluated whether the variants associated with PCOS in Han Chinese are associated with PCOS in white Europeans and concluded that DENND1A, THADA, FSHR, INSR and YAP1 loci are likely to play important roles in the etiology of PCOS across populations [11]. Polycystic ovary syndrome (PCOS), the commonest endocrine disorder affecting young women, appears to be a multigenic trait with contributing genes being unclear. Available genotyping methods are expensive, time-consuming with limited analytical sensitivity
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
