دراسة دور بعض العوامل في التغايرات الوراثية ونسبة الاصابة بفايروس COVID-19

رسالة ماجستير

اسم الباحث : سمر رفيع حربي خفيف

اسم المشرف : " الاستاذ المساعد زهير حميد عبود الاستاذ الدكتور حسن علي حسين السعدي

الكلمات المفتاحية :

الكلية : كلية العلوم

الاختصاص : علوم الحياة

سنة نشر البحث : 2024

تحميل الملف : اضغط هنا لتحميل البحث

الخلاصة

انتشرت متلازمة الجهاز التنفسي الحادة الناجمة عن فيروس كورونا بسرعة ، اذ غزت العالم وتسببت بوفاة أكثر من ستة مليون شخص. لذلك تهدف هذه الدراسة عزل وتشخيص أنواع الفطريات المصاحبة لـلخمج بفايروس كورونا وتحديد مسببات المرض وبعض الأعراض وبعض الأمراض المصاحبة للمرضى قيد الدراسة وكذلك تقدير مستوى بعض المعايير الدموية والمناعية وتحديد تعدد الأشكال الجينية لبعض الجينات.
شملت دراسة الحالية 167 (عينة دم ومسحة انفية) وزعت على النحو التالي: المجموعة الأولى كانت لـ 30 مريضًا مصابًا بـ Covid-19 وعدوى فطرية(17 ذكر,13 انثى) ، اما المجموعة الثانية كانت لـ107 مرضى مصابين بـ Covid-19 بدون عدوى فطرية (58 ذكر،49 انثى) في حين تمنت المجموعة الثالثة 30 شخصًا سليمًا كمجموعة سيطرة . وقد تراوحت اعمار عينات الدراسة بين10-70 عاما .
تم جمع العينات السريرية خلال الفترة (كانون الثاني 2022-نيسان 2022) من المرضى الراقدين في وحدات العناية المركزة كحالة خطيرة من مستشفى المرجان التعليمي (مدينة المرجان الطبية) ومستشفى الامام الصادق التعليمي بمحافظة بابل ومستشفى الزهراء بمحافظة كربلاء. زراعة العينات في SDA Agar و CHROM Agar واختبار انحلال الدم و تم تحديد مستويات بعض المعلمات الدموية والمناعية في الدم بواسطة ELISA و CBC … وباستخدام المعلومات في الاستبيان للكشف عن بعض الأعراض والأمراض المصاحبة بالاافة الى ذلك تمنت الدراسة الحالية التحري عن تعدد الاشكال النيوكليوتيد المفرد الى الجينات IL-17A and IRF-7 .
أظهر التحليل الإحصائي أن الذكور كانوا أكثر عرضة للإصابة من الإناث مع (p = 0.077) ، وتزداد الإصابة بـ COVID-19 مع تقدم العمر وأن كبار السن كانوا أكثر عرضة للاصابة من الشباب ، وكان هناك فرق كبير (p≤0.0001) بين ثلاث مجموعات في مستوى WBc ، العدلات ، الخلايا الليمفاوية ، Ferritin ، D-dimer ، CRP و suPAR وايضا هنالك فرق كبير (p≤0.0001) بين ثلاث مجموعات في وجود بعض الأعراض مثل الحمى والسعال وضيق التنفس وأعراض الجهاز الهضمي ووجد فرق معنوي بين ثلاث مجموعات في وجود بعض الأمراض المصاحبة.
أخيرًا ، اظهر التحليل الإحصائي للدراسة الجزيئية أن rs2275913 SNP للمركب الخلوي IL-17A يعتبر عامل خطر لـمجموعة COVID-19 مع العدوى الفطرية ، بينما لم يعتبر عامل خطر لـمجموعة COVID-19 بدون عدوى فطرية ، rs3819025 على IL-17A لم يعتبر الجين عامل خطر لـمجموعة COVID-19 مع العدوى الفطرية ، في حين يعتبر عامل خطر لـمجموعة COVID-19 مع العدوى الفطرية .
كذلك بين التحليل الاحصائي لتعدد الاشكال للجين RS 17155933في العامل المنظم للانترفيرون -7 انه لا يعتبر عامل خطورة بوجود او عدم وجود عدوى فطرية, في حين ان تعدد الاشكال الجيني للrs702965 في العامل المنظم للانترفيرون-7 يعتبر عامل خطر لمجموعة COVID-19بدون اصابات فطرية فقط ،اما تعدد الاشكال الجيني لل rs702966 للعامل المنظم للانترفيرون-7 فقد اعتبر عامل خطورة لمرضى COVID-19 في المجموعتين.
استنتجت نتائج الدراسة الحالية ان تعدد الاشكال الجيني للمركب الخلوي IL-17A والبروتين IRF-7 يكون مرتبط مع خطورة الاصابة بفايروس كورونا .
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Study The Role of Some Factors in Genetic Variations and The Rate of COVID-19 Virus

ABSTRACT

Acute respiratory syndrome caused by the Corona virus has spread rapidly, conquering the world and causing the death of more than six million people. Therefore, this study aims to isolate and diagnose the types of fungi associated with infection with the Corona virus, determine the causes of the disease, some symptoms, and some diseases associated with the patients under study, as well as estimate the level of some hematological and immunological parameters and determine the genetic polymorphisms of some genes.
The current study included 167 blood samples and nasal swabs(saliva) distributed as follows: The first group was for 30 patients with Covid-19 and a fungal infection (17 males, 13 females), while the second group was for 107 patients with Covid-19 without a fungal infection. (58 males, 49 females) while the third group wanted 30 healthy people as a control group. The ages of the study samples ranged between 10-70 years.
Clinical samples were collected during the period (January 2022 – April 2022) from patients hospitalized in intensive care units as serious cases from Al-Marjan Teaching Hospital, Imam Al-Sadiq Teaching Hospital in Babil Governorate, and Al-Zahra Hospital in Karbala Governorate. Culturing the samples on SDA Agar and CHROM Agar and testing for hemolysis. The levels of some hematological and immune parameters in the blood were determined by ELISA and CBC… and using the information in the questionnaire to detect some symptoms and associated diseases. In addition, the current study sought to investigate single nucleotide polymorphisms. To the genes IL-17A and IRF-7.
Statistical analysis showed that males were more susceptible to infection than females with (p = 0.077), COVID-19 infection increases with age and that older people were more susceptible to infection than young people, and there was a significant difference (p≤0.0001) between three groups in Level of WBc, neutrophils, lymphocytes, Ferritin, D-dimer, CRP and suPAR. There was also a significant difference (p≤0.0001) between three groups in the presence of some symptoms such as fever, cough, shortness of breath and gastrointestinal symptoms. A significant difference was found between three groups in the presence of some Comorbidities.
The statistical analysis of the molecular study showed that the rs2275913 SNP for the IL-17A cellular complex was considered a risk factor for the COVID-19 group with fungal infection, while it was not considered a risk factor for the COVID-19 group without fungal infection, and the rs3819025 gene for the IL-17A group was not considered a risk factor for the group. COVID-19 with fungal infection, while it is considered a risk factor for the COVID-19 with fungal infection group.
Also, the statistical analysis of the polymorphism of the gene RS 17155933 in the interferon-regulating factor-7 showed that it is not considered a risk factor in the presence or absence of a fungal infection, while the genetic polymorphism of rs702965 in the interferon-regulating factor-7 is considered a risk factor for the COVID-19 group without fungal infections only. As for the genetic polymorphism of rs702966 for the interferon-7 regulatory factor, it was considered a risk factor for COVID-19 patients in both groups.
We conclude from the results of the current study that genetic polymorphisms of the cellular complex IL-17A and the protein IRF-7 are associated with the risk of infection with the Coronavirus.

The acute respiratory syndrome caused by the Corona virus, which is one of the respiratory viruses that spread quickly, invaded the world and killed more than six million people. This study searched for several aspect including isolation and diagnosis of fungi species associated with COVID-19, and determined the pathogenic of it ,symptoms and some comorbidities in patients under study, so determined levels of some hematological and immune markers in blood of patients and detection polymorphisms in genes in specific SNPs and role this polymorphisms in immune of patients against diseases, compared this polymorphisms among groups. This case control study included 167 (blood specimens, nasal swaps) distributed as the following: first group included 30 patients with COVID-19 with fungal infection was 17 male and 13 female second group 107 patients with COVID-19 without fungal infection (after isolation and diagnosis of fungi in nasal swaps) was 58 male and 49 female ,Third group included 30 healthy persons as control group was 16 male and 14 female, All with ages (10-70) years .
The blood samples and nasopharyngeal swaps were employed for the detection whether the persons infected by SARS-CoV-2 by SARS-CoV-2 Antigen Rapid Test Kit, that proceeded by collected the specimens between January-2022 to April – 2022, Clinical samples were collected from patients whom lying in Restorative Care Unit as severe case from Marjan Teaching Hospital , AL-Imam AL-Sadiq Teaching Hospital in Babylon province and AL-zahraa Hospital in Karbala Province. After cultivation of specimens in SDA agar ,CHROM agar and hemolysis test, determined levels of some hematological and immune markers in blood by ELISA, CBC,…, By using of information in questionnaire to detection some symptoms and comorbidities finally molecular study .Statistical analysis showed that male ware more prevalence to disease than female with p=0.077, the elderly people were more affected with COVID-19 than young people, there were significant difference with p ≤0.0001, among three groups in level of WBC, Neutrophils, Lymphocyte, Ferritin, D-dimer, CRP and suPAR this is the first study in Iraq that studied the level of suPAR in the blood of COVID-19 patients, high significant difference with p ≤0.0001among three groups in presence of some symptoms like fever, cough, dyspnea and gastrointestinal tract GIT symptoms, Significant difference among three groups in the presence of some comorbidities.
Finally, the molecular study statistical analysis found that rs2275913 SNP on the IL-17A gene was considered a risk factor for COVID-19 with fungal infections patients, while did not considered as a risk factor for COVID-19 without fungal infections group, rs3819025 on IL-17A gene did not considered a risk factor for COVID-19 with fungal infections patients, but considered as a risk factor for COVID-19 without fungal infections group, rs17155933 on IRF7 gene did not considered a risk factor in two groups, while rs702965 on IRF7 considered as a risk factor for COVID-19 without fungal infections group this is the first study in Iraq and the world which associated polymorphisms in these two SNPs with COVID-19 patients , rs702966 SNP on the IRF-7 gene and considered as a risk factor for COVID-19 patients in two groups, Polymorphism in IL-17 A and IRF-7 were associated with the severity of COVID-19 patients.