A novel tetra-primer ARMS-PCR for genotyping of the OPRM1 gene rs1799971 variant associated with opioid use disorders

• Published in: BMC research notes 2023-11, Vol.16 (1), p.333-333, Article 333
• Main Authors: Wijekumar, P J, Ranadeva, N D K, Jayamaha, A R, Herath, H M N D M, Noorden, N, Fernando, S S N
• Format: Article
• Language: English
• Subjects: Addictions; Alleles; Annealing; DNA microarrays; Drug abuse; Drug addiction; Gene Frequency; Genetic diversity; Genetic testing; Genomes; Genotype; Genotype & phenotype; Genotypes; Genotyping; Humans; Mass spectroscopy; Morphine; Narcotics; Opioid use disorder; Opioid-Related Disorders - genetics; Opioids; OPRM1; Optimization; Polymerase Chain Reaction; Polymorphism, Single Nucleotide; Receptors, Opioid, mu - genetics; rs1799971; Software; T-ARMS PCR; Temperature
• ISSN: 1756-0500; 1756-0500
• DOI: 10.1186/s13104-023-06578-7

A SNV is a single nucleotide change that can occur at any point in the genome. SNVs are the most common genetic variants that occur in the human genome, and a number of SNVs have been found to be associated with human traits and disease. Researchers genotype SNVs using TaqMan technology, DNA microarray, MALDI-TOF mass spectrometry, and automated sequencing, which are expensive and time-consuming. The OPRM1 gene rs1799971 (A118G) has been identified for its association with Opioid use disorder (OUD). The present study focused on developing a single step identification test using Tetra-Primer Amplification Refractory Mutation System-PCR (T-ARMS-PCR) to detect the presence of SNV OPRM1 rs1799971 (A118G). This study was performed to optimize the protocol for the designed four primers and validate it using a total of 52 buccal samples from volunteers who are currently under rehabilitation for the drug abuse disorder. Utilizing 52 DNA samples, a novel T-ARMS-PCR assay was successfully developed, tested, and validated. The products of the T-ARMS PCR for rs1799971 contained 395 bp as the control band, 186 bp as G allele (variant) and 257 bp as A allele (wild type), which 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 G allele frequency was 56.7% and A allele frequency was 43.3%.