رسائل ماجيستير
English
2022
Study of Human Gene Polymorphism Associated with Covid-19 Infection in Type 2 Diabetes Mellitus Patients
علي حمود السعدي, مريم علي حسين علوان
كلية العلوم-جامعة بابل · العراق
Compared with previous epidemic infections, Coronavirus has the highest transmission rate and risk of death. On the other hand, diabetes is one of the most prevalent diseases. Thus, diabetics infected with the COVID-19 virus are the most affected. The aim of this research was to find out if there was a correlation between infection with the COVID-19 virus and patients with type 2 diabetes. The study concluded that there has been a relationship between diabetes and patients with COVID-19. The current study was conducted in the molecular laboratory in the Department of Biology, College of Science, University of Babylon. The samples were collected, and the practical part of this study was completed during October (2021-2022). The samples were obtained from patients with COVID-19 and COVID-19, diabetes mellitus, and healthy people. These samples were collected from Marjan Medical Hospital and Al-Saadiq Teaching Hospital in Babylon Governorate, where the total number of samples was 92, including 31 blood samples from healthy people (3 females, 28 males), 30 blood samples from patients infected with COVID-19 (16 females, 14 males), and 31 blood samples from patients infected with COVID-19 + diabetes (17 females, 14 males).
The current study has been divided into three parts: the first study was demographic, the second included measuring blood parameters, and the third included a molecular study. The first part included the social and demographic distribution of the study coefficients in terms of gender, level of injury, age, duration of injury, and body mass. The second part included an assessment encompassing the estimation of physiological parameters (HBA1C, FBG, insulin, insulin resistance, insulin sensitivity, CRP, D-dimer, Ferritin), and the third part involved the study of the effects of some genes (ACE2, AT1R, rs657152) determined by PCR.
The results of this study showed that gender revealed significant differences at the probability value (p≤0.01) (0.0002) between the two groups of males and females, where the males in the CDM2 group were (46.67%) while in the C group it was (43.3%). The value of females was (56.7%) in group C and it was (48.38%) in group CDM2. The level of infection was classified into three sub-levels including: mild, moderate, and severe. The percentage of the level of cut infection in CDM2 (48.30%) was higher than that of group C (26.66%), while the average level was higher in CDM2 group (38.70%) than it was before. Therefore, the minor level was lower in CDM2 (12.9%) than in group C (43.33%), and the severity of infection increased in CDM2 than in group C.
As for the age groups, there were statistically significant differences for the three categories adopted in the current study (>30, 30-50, <50 years). The group of less than 30 years old was more in group C (36.7%) than in the CDM2 category (6.5%), while the 30-50 category was rounded off in the C and CDM2 groups (26.7%, 22.6%). A higher percentage of greater than 50 categories was observed in CDM2 (71%) and the total of C was (36.7%). There were three BMI categories according to the current study including (normal, overweight, and obesity), and there were no statistically significant differences among these groups. As for the injury period, the duration periods were classified into 3 subgroups including (<7, 7-14, >14 days). It was less than 7 days in group C (73.3%) while it was (51.61%) in CDM2, the 7-14 days in group C was (13.33%) and it was (32.25%) in CDM2 group, and more than 14 days was (13.33%) in the CDM2 group and it was (16.12%) in the C group. The differences among groups were significant (p. 0.0311) with a value of (p ≤ 0.05).
The results of the physiological study showed significant differences among healthy people and COVID patients along with patients with COVID + diabetes mellitus compared to healthy people in ferritin (108.59±18.4, 322.28±30.1, and 583.10±44.6) respectively, while significant differences at (p≤0.01), showed significant differences in FBG, cumulative glucose, and insulin resistance only in the COVID-19 + diabetes group (253.09±16.2, 9.15±2.0, and 17.51±4.1 respectively) at (p≤0.05) (p≤0.01), and at the same time there was no significant difference among the healthy groups. C-reactive protein or D-dimer showed high values in the COVID-19 group and the COVID-19 + diabetes groups compared with the healthy groups (43.72±5.3, 61.18±5.3, 1731.41±114.5, and 1282.97±221.3) at (p≤0.01) respectively, but it did not show significant differences in insulin and insulin sensitivity.
In the molecular study, single nucleotide polymorphism (SNPs) of ACE2 gene was detected by using PCR technique in patients compared with the healthy groups. The group (DD; ID) showed statistically significant differences in relation to the genetic makeup of patients compared with healthy controls at (0.04), whereas I, II did not show any significant relationship. The ACE D genotype was dominant in three groups: (72.6%, 85%, 54%) in the healthy control group, the COVID-19 group, and the COVID-19 + diabetes group respectively, followed by the alleles DD; ID; I. It was observed in AT1R that there was a very significant difference between the healthy group and the group of patients by using polymerase chain reaction techniques PCR-SSCP, where the distribution of DNA polymorphisms was found to be (96.77%, 3.23%) in the healthy group and (6.67%, 93.33%), respectively. In COVID-19 + diabetes group and (12.9%, 87.1) and in COVID-19 + diabetes patients, the results showed that there was no association between DNA polymorphisms according to the number of bands with patients compared to control groups. Meanwhile, there were significant differences in frequencies of the AT1R polymorphism among patients with COVID-19 and COVID-19 with diabetes mellitus and healthy subjects at (P-value ≤ 0.01).
As for the rs567152 SNP, the study showed that there were no statistically significant differences among the genetic makeup of patients compared with the genetic makeup of healthy people. Finally, it was observed that patients with diabetes who contracted COVID-19 appeared to have significantly higher levels of (CRP, D-dimer, and Ferritin) and parameters related to diabetes than patients who suffered from COVID-19 alone, also high levels of ACE2 gene, AT1R receptor as such this would make them more likely to develop illness and death.