Genotyping of alleles causing alpha-1 antitrypsin deficiency (A1ATD) in SERPINA1 gene by ARMS technique

Alpha-1-antitrypsin deficiency (AATD) is an autosomal co-dominant disease characterized by low-than-normal serum AAT levels caused by mutations in the gene called SERPINA1. AAT is produced in hepatocytes and released into the blood, and its primary function is to protect the body from neutrophil elastase activity (antiprotease). Deficiency in serum AAT level can cause damage to lung tissue by neutrophil elastase and other proteolytic enzymes, and consequently diseases such as emphysema, chronic bronchitis, chronic obstructive pulmonary disease (COPD), bronchiectasis and asthma. More than 100 different mutations have been identified in the SERPINA1 gene to date. Among these, the most common variants associated with the disease were identified as S and Z alleles. Normal AAT protease inhibitor (PI) is denoted by the letter 'M'. Methods such as immunoassay methods, isoelectric focusing (IEF) and molecular genotyping (PCR and PCR-RFLP) are used to detect variants. The amplification-refractory mutation system (ARMS) is a simple, fast and reliable method to detect any mutation containing single base changes or small deletions. When we look at the literature, there is no genotyping study using the ARMS method in patients with COPD and Emphysema. In this planned study, it was aimed to genotype the most commonly associated 'S' and 'Z' variants and their normal alleles with the ARMS-PCR method. Peripheral blood was taken from the patient group (53 individuals) diagnosed with COPD and emphysema and the control group (33 individuals), their DNA was isolated, and PCR amplifications were performed with the designed specific primer sequences. Sequence analysis of the relevant regions of all samples was performed to confirm the ARMS-PCR results. In conclusion; With this method, normal allele and 'S' variant could be determined but 'Z' variant and normal allele could not be detected specifically despite using three different primers.