A Novel Tetra-Primer ARMS-PCR for genotyping of the OPRM 1 gene rs1779791 variant associated with opioid use disorders

Objectives: SNV is a single nucleotide change that can occur at any point in the genome. It is regarded as a frequent genetic variant that can result in genetic diseases. Discovering genetic variants in individuals is one of the most essential jobs of genetic research. Researchers genotype SNVs using TaqMan technology, DNA microarray, MALDI-TOF mass spectrometry, and automated sequencing which is expensive and time-consuming. The OPRM1 gene rs1799971 (A118G) has been identified for its association with Opioid use disorder (OUD). The present study focused in developing a single step identification test using Tetra-Primer Amplification Refractory Mutation System-PCR (T-ARMS-PCR) in order to detect the presence of SNV OPRM1 rs1799971 (A118G). The present research was performed to optimize the protocol for the designed four primers and validate using a total of 52 buccal samples from volunteers who are currently under rehabilitation for the drug abuse disorder. Results: The novel assay was successfully designed, tested, and validated using 52 DNA samples. The products of the T-ARMS PCR for rs1799971 contained 395bp as the control band, 186bp as G allele (variant) and 257bp as A allele (wild type) were observed in the gel image. The genotype frequencies for the OPRM1 gene rs1799971 (A118G) were 44% (22/52) of homozygous variant type (GG), 28.9% (15/52) of homozygous wild type (AA) and 28.9% (15/22) of heterozygous (AG). The minor allele (G) frequency was 56.7% and major allele (A) frequency was 43.3%.