Detection of ACE Gene SNPs Using rhAmp Genotyping Platform and Their Association with I/D Polymorphism in COVID-19 Patients with Hypertension

Abstract

The high activity of Angiotensin Converting Enzyme (ACE) in patients with hypertension (HT) potentially affects the risk and severity of COVID-19 infection. The ACE gene variation leads to the modification of ACE activity in the Renin-Angiotensin System (RAS). Therefore, this study aims to identify three ACE gene SNPs using the rhAmp SNP genotyping platform, i.e., rs4331 (A/G), rs4341 (G/C), and rs4343 (G/A), as well the linkage disequilibrium (LD) with rs1799752 (I/D) polymorphism. It also analyzed their associations with COVID-19 severity and comorbid HT. This cross-sectional study was carried out in two regions of Indonesia, i.e., Palu City, Sulawesi, and Lahat District, Sumatera, each representing the eastern and western parts, with a total of 95 COVID-19 patients recruited in 2021. The SNP detection results showed that the percentage of genotypes GG, AG, and AA, i.e., 57%, 32%, and 11%, respectively, at rs4331, corresponded with II, DI, and DD at rs1799752. However, at rs4341, only genotype GG was found, while at rs4343, 37% of the samples were undetermined. The results also revealed that rs4331, rs1799752, and rs4343 showed strong LD (D'= 0.97 and 0.99), while rs4331 and rs1799752 were considered replaceable and can serve as gene markers (R2= 0.957). Genotype rs4343 and haplotype analysis showed a significant difference between the residents of western and eastern Indonesia (p< 0.05), while no difference was observed for comorbid HT and the severity of COVID-19. About 17% of the G-I-G-G haplotypes indicated moderate-severe (M-S) severity, while 15% of the A-D-G-G haplotypes showed mild (M) severity. It was concluded that the rs4331, rs4343, and rs4341 genotypes could successfully be detected by using the rhAmp genotyping platform. However, the association with I/D polymorphism was likely co-inherited for rs4331 and rs4343 only. The two dominant haplotypes associated with mild and moderate-severe symptoms were A-D-G-G and G-I-G-G, respectively.