بررسی جهش های پروتئین نوکلئوکپسید (N) ویروس کرونا در دامنه موثر در قدرت ایمنی زایی و تاثیر آن بر تست های تشخیص سرولوژیک در نمونه های جدا شده از بیماران مبتلا در استان زنجان
محورهای موضوعی : تشخیص مولکولی نشانگر های بیوشیمیایی و ژنتیکی
سمانه کریم خانی لویی
1
(گروه ژنتیک مولکولی، واحد اهر، دانشگاه آزاد اسلامی، اهر، ایران.)
سعید قربیان
2
(گروه ژنتیک مولکولی، واحد اهر، دانشگاه آزاد اسلامی ، اهر ، ایران)
ساناز مهمازی
3
(گروه زیست شناسی، دانشکده علوم پایه ، واحد زنجان، دانشگاه آزاد اسلامی، زنجان، ایران)
چنگیز احمدی زاده
4
(گروه میکروبیولوژی، واحد اهر، دانشگاه ازاد اسلامی، اهر، ایران)
کیوان ندائی
5
(گروه بیوتکنولوزی پزشکی، دانشکده پزشکی ، دانشگاه علوم پزشکی زنجان، زنجان، ایران.)
کلید واژه: نوکلئوکپسید, کووید-19, پروتئین N, جهش, IgG,
چکیده مقاله :
زمینه و هدف: کووید-19 یک بیماری ویروسی با شیوع جهانی است که به دلیل جهش¬های مختلفی که در ژنوم ویروس رخ میدهد، دارای واریانت¬های متعددی بوده و شدت و تظاهرات بالینی متنوعی دارد. یکی از پروتئینهای مهم در پاتوژنز و ایمنی زایی ویروس، پروتئین نوکلئوکپسید یا پروتئین N است که دومینهای مختلفی دارد و ایمونوژن ترین دومین آن جهت ارزیابی جهشهای موجود در این تحقیق مورد بررسی قرار گرفت. مواد و روش¬ها: در این مطالعه در مجموع از 85 بیمار مبتلا به کرونا در استان زنجان نمونه برداری صورت گرفت، و سپس از این نمونهها RNA ویروسی استخراج شده و cDNA ساخته شد. سپس با انجام PCR برای نقاط Hot Spot پروتئینN محصول PCR توالی یابی گردید. توالیها از نظر وجود جهش مورد غربالگری قرار گرفته و فاکتورهای سرمی IgG و IgM مربوط به پروتئین N به روش الیزا مورد ارزیابی قرار گرفت. نتایج: جهش¬های 28881-2-3GGG>AAC، C28977T،C28932T ، C28854T، G28975T، G28881T، C28830T، C28905T و C28977T در پروتئین N مشاهده شدند، که این جهش¬ها در تشخیص ویروس با روش¬های مولکولی تداخل ایجاد می¬کنند. میزان آنتی بادی¬های سرمی IgM و IgGبا تست الایزا در بیماران بررسی شد. بیشترین میزان جهش در نوکلئوتید C28854T بود که منجر به تبدیل S194L میشد. همچنین این موارد جهش یافته تیتر IgG بالاتری داشتند. یک مورد جهش C28830T مشاهده شده در نمونهها از جهش های شناسایی شده در نمونههای ایران بوده است، که منجر به تغییر S186F شده است. نتیجه گیری: منطقه مورد بررسی شامل بخشی از ژنوم ویروس بود که دامنه اتصال به RNA ویروسی پروتئین N را شامل میشد. این دومین قدرت ایمنی زایی بالایی داشته و میتواند در طراحی کیتهای تشخیص ایمونولوژیک سریع، همچنین در پروب کیتهای تشخیص مولکولی مورد استفاده قرار گیرد. این پروتئین که توالی آن نسبتا حفاظت شده است، در واریتههای مختلف ویروس به علت اهمیت عملکردی و با توجه به جهشهای اختصاصی میتواند در طراحی واکسنها با در نظر گرفتن انواع ساب تایپها مورد استفاده قرار گیرد.
Background &Aim: Wellness requires the proper functioning of the immune system in all of the body systems. The immune system active by the cooperation of several immune sections. The immune cells and substances can identify and destroy foreign viral and microbial agents that worn out cells and cancer cells in the body. The aim of this study was to investigate the effect of high-speed sports activity on the anti-inflammatory factors lysozyme, LL-37 and HDB-2 in the saliva of obese adolescent boys. Materials &Methods: 32 male students voluntarily participated in the present study and were randomly assigned to four groups: 1) obese aerobic exercise, 2) normal weight aerobic exercise, 3) obese control without exercise, and 4) normal weight control without exercise. Anthropometric indices of height, weight and body mass index were measured. Before and after eight weeks of practice, Shatell-Run standard test was performed. Salivary samples of lysozyme, lactoferrin, lactate and C-reactive protein concentrations were taken after eight weeks of training with a frequency of three sessions per week with an intensity of 30 to 90% of maximum aerobic power. Using analysis of covariance, variables with a significance level of less than (p≥0.05) were included in the analysis. Results: The results showed that the salivary levels of lysozyme (p=0.001), LL-37 (p=0.002), and HDB2 (p=0.001) increased significantly, and the amount of increase in obese students was higher than in people with normal weight. Conclusion: The increased response of some salivary anti-inflammatory proteins following eight weeks of high-speed exercise training after increased activity may be due to the short-term responses of the immune system against the pressures caused by intense activity.
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