تکثیر هم‌دمایی به واسطه حلقه: روش تشخیص سریع و کم هزینه عوامل عفونی
الموضوعات :
اعظم عسکری
1
,
محمد کارگر
2
,
صادق قربانی دالینی
3
,
عباس دوستی
4
1 - دانشگاه آزاد اسلامی، واحد جهرم، گروه میکروبیولوژی
2 - دانشگاه آزاد اسلامی، واحد جهرم، گروه میکروبیولوژی
3 - دانشگاه آزاد اسلامی، واحد جهرم، باشگاه پژوهشگران جوان
4 - دانشگاه آزاد اسلامی، واحد شهرکرد، مرکز تحقیقات بیوتکنولوژی
الکلمات المفتاحية: بهینه سازی, LAMP, طراحی پرایمر,
ملخص المقالة :
روش تکثیر هم دمایی به واسطه حلقه (LAMP) مبتنی بر تکثیر اسید نوکلئیک در دمای ثابت ˚C 65-62 است. در این روش بدون نیاز به مرحله واسرشت شدن حرارتی، به کمک آنزیم DNA پلی مراز Bst به صورت هم زمان امکان جداسازی و تکثیر دو رشته DNA وجود دارد. بنابراین بر خلاف سایر روش های متداول تکثیری ، بدون نیاز به چرخه های دمایی و دستگاه ترموسایکلر و با استفاده از تجهیزات ارزان قیمتی مانند حمام آب گرم و یا بلوک حرارتی قابل انجام است. برای طراحی واکنش LAMP نیاز به 6 عدد پرایمر وجود دارد که توانایی اتصال کاملاً اختصاصی به 8 ناحیه مجزا از توالی هدف را داشته باشند. در نتیجه واکنش LAMP مقدار بسیار زیادی محصول ایجاد می شود که بدون نیاز به روش پر زحمت الکتروفورز در ژل، به کمک روش های ساده تری مانند مشاهده کدورت و یا تغییر رنگ ناشی از رنگ های فلورسنس در مخلوط واکنش به سادگی قابل تشخیص می باشد. بدین ترتیب روش LAMP با توجه به مزایایی مانند حساسیت و ویژگی مناسب ، کارایی بالا ، عدم نیاز به تجهیزات آزمایشگاهی گران قیمت و سادگی تشخیص، می تواند به عنوان ابزاری ارزشمند برای تشخیص سریع بیماری های عفونی در آزمایشگاه های کلینیکی و بیمارستانی به ویژه در کشورهای درحال توسعه به کار برده شود. این مقاله با هدف معرفی مبانی و کاربردهای روش LAMP در تشخیص عوامل عفونی تدوین گردیده است.
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