Immunological and Allele Specific Genotyping Study Among Patients with SARS-CoV-2
زيتون عبد الرضا الخفاجي, فلاح حسن عبيس جبر
كلية الطب-جامعة بابل · العراق
The disease caused by SARS-CoV-2 known as COVID-19, was considered a pandemic by the World Health Organization (WHO) and had the characteristic of being highly contagious and of rapid spread, which led to changes in human habits with an impacted the global health The current study included 125 patients and 60 enrolled as a control group for immunological and genotyping investigations. All patients were admitted to Marjan Medical City, Al-Shomali general hospital, and Al-Sadeq hospital’s COVID‑19 ward in the period between January 2022 to July 2022. The diagnosis COVID‑19 in each patient was confirmed by SARS-CoV-2-positive RT-PCR. Single nucleotide polymorphism (SNPs) for IL-12A-rs568408 and TYK2-rs2304256 were detected by Allele specific-PCR method.ELISA test used to determine concentration of interleukin-35 (IL-35)and presepsin(PSN)in serum. Patients’ group included 56 (44.8%) males and 69 (55.2%) females, whereas, control group included 28 (46.6%)males and 32 (53.4%) females and there was no significant difference in the frequency distribution of patients and control subjects according to sex(P = 0.87).As well as there was no significant difference regarding age between patients and control subjects (P = 0.65).IL-35 showed statistical significant differences between patients 6.86± 2.31ng/ml and control group 3.86± 2.07 (ng/ml) (P< 0.0001). PSN showed statistical significant differences between patients 318.18± 226.62 (pg/ml) and control group 199.68± 39 (pg/ml) (P< 0.0001). The distribution of both genotyping and allele frequencies of IL-12A rs568408 revealed significant differences between patients and control groups (P= 0.006 and p=0.001 respectively). The IL12A rs568408 AA and AG variant genotypes were associated with a significantly increased risk of COVID-19[odds ratio (OR) = 5.19, 95%Confidence interval (CI): 1.13-23.82; P= 0.034] and [OR = 2.39, 95% CI = 1.16-4.94, P= 0.018] respectively compared with the wild-type GG homozygote.Logistic regression analysis revealed that the frequencies of the homozygous variant AA and heterozygous variant AC of TYK2-rs2304256 were 14.4 % and 22.4 % in COVID-19cases and 3.3 % and 16.7 % in healthy controls, respectively. For AA genotype (OR= 5.46, 95% CI: 1.21-24.61; P= 0.027). While AC genotype (OR = 1.7, 95% CI = 0.76-3.81, P = 0.196), at TYK2-rs2304256 demonstrated that AA genotype showed a statistically significant risk for COVID-19in Iraq compared with the wild-type rs2304256 CC. The variant rs568408 AC/AA genotypes were associated with a significantly increased risk of COVID-19(OR= 2.81, 95% CI: 1.37-5.78; P= 0.004), compared with the wild-type rs2304256 CC.In conclusion, IL-35 and Presepsin showed statistical significant differences between patients and controls (P< 0.0001) for each one. IL-35 can be used in diagnosis of COVID-19. The IL12A rs568408 AA and AG variant genotypes were associated with a significantly increased risk of COVID-19. In TYK2-rs2304256, AA genotype showed a statistically significant risk for COVID-19 compared with the wild-type rs2304256 CC