Development of amplification refractory mutation system polymerase chain reaction-based assay for genotyping the single nucleotide polymorphisms of MED12 gene: An economical approach for case–control investigations of uterine leiomyoma

Uterine leiomyomas (UL), commonly known as fibroids, are non-cancerous tumors that develop in the uterine myometrium within the female reproductive system. Investigating single nucleotide polymorphisms, particularly mutations, is vital in understanding cancer and other diseases, assisting in genotypic classification, and guiding treatment and genetic counseling. This study aimed to develop and optimize the detection of the MED12 gene (rs199469672) G/A polymorphism in patients with ULs. A total of 160 participants were included in the study, where the tetra-primer amplification refractory mutation system polymerase chain reaction (ARMS-PCR) method was used to assess allele and genotype frequencies. The distribution of genotypes for the MED12 gene (rs199469672) G/A polymorphism was as follows: Homozygous GG at 37.7%, heterozygous GA at 56.3%, and homozygous AA at 6.2%. The allele frequencies were 65.6% for G and 34.4% for A. The genotype distribution conformed to Hardy–Weinberg equilibrium, with a P -value of 0.18. In conclusion, the somatic mutation (rs199469672) in MED12 appears to be tissue-specific, likely occurring in certain cell subsets within specific tissues. The absence of this mutation in blood samples reinforces the idea that it is a somatic event, possibly associated with localized conditions such as tumors or tissue-specific diseases. To further explore this mutation and its potential role in disease, additional studies are necessary, including larger and more varied tissue samples, as well as advanced techniques for isolating and detecting somatic mutations in blood, such as circulating tumor deoxyribo nucleic acid (DNA). This study demonstrates a straightforward, fast, and cost-efficient ARMS-PCR method for detecting allele-specific DNA polymorphisms and mutations.