ساخت زیست حسگر الکتروشیمیایی بر مبنی DNA فاقد نشانگر و اصلاح شده با نانو مواد جهت شناسایی ویروس موزاییک انجیر
نیلوفر رجبی
1
,
محمد رضا صفرنژاد
2
,
فرشاد رخشنده رو
3
,
مسعود شمس بخش
4
,
حجت الله ربانی
5
1 - گروه گیاهپزشکی، دانشکده علوم کشاورزی و صنایع غذایی، واحد علوم و تحقیقات دانشگاه آزاد اسلامی، تهران. صندوق پستی: 775- 14515 – کدپستی: 147789385
2 - بخش تحقیقات ویروس شناسی، موسسه تحقیقات گیاه پزشکی کشور، سازمان تحقیقات آموزش و ترویج کشاورزی
3 - گیاه پزشکی، دانشکده علوم کشاورزی و صنایع غذایی، واحد علوم و تحقیقات دانشگاه اسلامی، تهران
4 - استاد گروه بیماری شناسی گیاهی، دانشکده کشاورزی، دانشگاه تربیت مدرس
5 - گروه مهندسی آنتی ژن و آنتی بادی مرکز تحقیقات آنتی بادی مونوکلونال پژوهشکده ابن سینا، دانشگاه شهید بهشتی، تهران، ایران
کلید واژه: ویروس موزاییک انجیر, زیست‌حسگر الکترو شیمیایی, بهینه¬سازی, کالیبراسیون,
چکیده مقاله :
امروزه زیست¬حسگر¬های الکترو¬شیمیایی به¬دلیل حساسیت و دقت بالا در ردیابی اسید¬های نوکلئیک هدف مورد توجه بسیار قرار گرفته¬اند. از طرفی به دلیل اهمیت اقتصادی درختان انجیر، تشخیص سریع و دقیق بیماری¬های آن از جمله بیماری موزائیک انجیر، Fig Mosaic Disease (FMD)، به¬عنوان یکی از مهم¬ترین و مخرب¬ترین بیماری¬های ویروسی درختان انجیر، نقش مؤثری در مدیریت این بیماری ایفا می¬کند. در این مطالعه، نانو¬زیست¬حسگر الکترو-شیمیایی مبتنی بر DNA و فاقد نشانگر مولکولی برای شناسایی کیفی و کمی ویروس FMV در نمونه‌های آلوده بهینه¬سازی شد. در این راستا، جهت بهینه¬سازی و بهبود عملکرد زیست‌حسگر طراحی شده، پارامتر¬های غلظت کاوشگر DNA برای تشکیل تک‌لایه‌های خود انباشته و زمان لازم جهت انجام فرآیند هیبریداسیون در محدوده زیستی با بیشترین تأثیر در زیست‌حسگر DNA، مورد بررسی قرار گرفت. با توجه به داده‌های منحنی کالیبراسیون، اولین محدوده خطی 3- میکرومولار تا 3 میکرومولار با معادله خط y = 4.81x+34.58 و ضریب تشخیص (9967/0=R2) به-دست آمد. زیست¬حسگر بهینه¬سازی شده در این پژوهش جهت تشخیص ماده ژنتیکی FMV دارای پایداری،گزینش‌پذیری بالا و همچنین کاربری آسان و سریع بوده که نوید یک روش تشخیصی با کارایی و دقت بالا جهت شناسایی این ویروس را می¬دهد.
Today, electrochemical biosensors are highly regarded due to their high sensitivity and accuracy in tracking target nucleic acids. On the other hand, due to the economic importance of fig trees, rapid and accurate diagnosis of its diseases, including Fig Mosaic Disease (FMD), which is one of the most important and destructive viral diseases of fig trees plays an effective role in the management and control of this disease. In this study, a novel label free electrochemical DNA-based biosensor was optimized to qualitatively and quantitatively identify the presence of FMV in the infected samples. In this regard, in order to optimize and improve the performance of the designed biosensor, the concentration parameters of the DNA probe to form self-assembled monolayers and the time required to perform the hybridization process in the biological range with the greatest impact on the DNA biosensor were investigated. According to the data of the calibration curve, the first linear range at -3 μM to 3 μM was obtained with the equation of the line y= 4.81x+34.58 and the detection coefficient (R2= 0.9967). In this research, the omtimized biosensor to detect a stable, high selective, easy and fast FMV-DNA, promises a diagnostic method with high efficiency and accuracy.
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