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:: دوره 24، شماره 5 - ( دو ماهنامه طب جنوب 1400 ) ::
جلد 24 شماره 5 صفحات 480-469 برگشت به فهرست نسخه ها
طراحی و ارزیابی عملکرد PCR جهت تشخیص ویروس سندروم لکه سفید در میگو
حسین معین ، فرشید کفیل‌زاده 1، محمدرضا میری، محمد خلیل‌پذیر
1- ، f.kafilzadeh@gmail.com
چکیده:   (186 مشاهده)
زمینه: ویروس سندروم لکه سفید، عامل بیماری لکه سفید در میگوها و بسیاری از سخت‌پوستان می‌باشد. این بیماری بسیار مسری است و در طی 3-10 روز در شرایط کشت طبیعی باعث مرگ‌و میر می‌شود، بنابراین، تشخیص زود هنگام این ویروس یک ضرورت می‌باشد.
مواد و روش‌ها: برای 3 ناحیه از ژنوم ویروس و یک ناحیه از ژنوم میگو پرایمرهایی سازگار با هم طراحی شدند که بتوانند با هم در یک واکنش عملکرد داشته باشند.استخراج DNA از 40 نمونه میگوی مشکوک به بیماری انجام گردید. سپس پرایمرها به صورت تکی جهت تشخیص ویروس لکه سفید بهینه‌سازی شدند و پس از انتخاب مناسب‌ترین آن‌ها، تشخبص ویروس به صورت همزمان توسط دو جفت پرایمر انجام گردید. از پرایمرهای تکثیر کننده ژنوم میگو هم به‌عنوان کنترل داخلی استفاده شد.
یافته‌ها: از بین پرایمرهای طراحی شده 3 جفت پرایمر که یک قطعه از ژنوم میگو و 2 قطعه از ژنوم ویروس را تکثیر می‌کردند انتخاب گردید. از 40 نمونه بررسی شده 28 نمونه مثبت (آلوده به ویروس) و 12 نمونه منفی بود که با نتایج به‌دست آمده با کیت مرجع مطابقت داشت.
نتیجه‌گیری: جهت تشخیص ویروس، بررسی 2 ناحیه از ژنوم آن کافی می‌باشد و احتمال منفی کاذب را کاهش می‌دهد. همچنین استفاده از ژنوم میگو جهت کنترل مراحل استخراج DNA و PCR مفید می‌باشد. روش PCR تک مرحله‌ای نسبت به روش‌های دو مرحله‌ای (Nested PCR) بدلیل کاهش احتمال آلودگی و راحتی انجام آن اولویت دارد.
واژه‌های کلیدی: ویروس، سندروم لکه سفید، PCR، تشخیص
متن کامل [PDF 467 kb]   (70 دریافت)    
نوع مطالعه: پژوهشي | موضوع مقاله: میکروب‌شناسی و ایمنولوژی
دریافت: 1400/9/7 | پذیرش: 1400/9/7 | انتشار: 1400/9/7
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Moein H, Kafilzadeh F, Miri M R, Khalilpazir M. Design and Evaluation of a PCR Method for Detecting White Spot Syndrome Virus in Shrimp. Iran South Med J. 2021; 24 (5) :469-480
URL: http://ismj.bpums.ac.ir/article-1-1516-fa.html

معین حسین، کفیل‌زاده فرشید، میری محمدرضا، خلیل‌پذیر محمد. طراحی و ارزیابی عملکرد PCR جهت تشخیص ویروس سندروم لکه سفید در میگو. طب جنوب. 1400; 24 (5) :480-469

URL: http://ismj.bpums.ac.ir/article-1-1516-fa.html



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