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A reverse transcriptase-loop mediated isothermal amplification assay (RT-LAMP) for rapid detection of bovine viral diarrhea virus 1 and 2 | ||
| Archives of Razi Institute | ||
| مقاله 1، دوره 72، شماره 2، مهر 2017، صفحه 73-81 اصل مقاله (512.3 K) | ||
| نوع مقاله: Original Articles | ||
| شناسه دیجیتال (DOI): 10.22092/ari.2017.109836 | ||
| نویسندگان | ||
| Amir Tajbakhsh1؛ Elham Rezatofighi* 1؛ Khalil Mirzadeh2؛ Mahdi Pourmahdi3 | ||
| 1Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran | ||
| 2Department of Animal Science and Food, Khouzastan Ramin Agriculture and Natural Resources University, Ahvaz, Iran | ||
| 3Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran | ||
| چکیده | ||
| Bovine viral diarrhea virus (BVDV) is a pathogen that infects cattle, and is globally important. It causes substantial financial losses to the livestock industry. In the current study, a one-step reverse transcriptase-loop-mediated isothermal amplification (RT-LAMP) assay was set up for rapid and efficient detection of BVDV. For this purpose, four primers were designed to recognize six distinct regions on the target RNA based on a highly conserved sequence in the 5΄ UTR of the BVDV genome. Eighty blood specimens were collected from bovines suspected to suffer from BVDV infection, and were tested in parallel by RT-LAMP and RT-PCR. Twenty four of these samples were positive by RT-LAMP, while twenty were positive by RT-PCR. The RT-LAMP detection limit was estimated to be approximately 70PFU /mL of virus. Comparison of RT-PCR with RT-LAMP in this study revealed the recent developed RT-LAMP a highly sensitive and specific for BVD virus detection in the clinical samples. | ||
| کلیدواژه‌ها | ||
| Bovine viral diarrhea؛ Reverse Transcriptase-Loop mediated isothermal amplification assay؛ Reverse Transcriptase-Polymerase Chain Reaction | ||
| عنوان مقاله [English] | ||
| طراحی و ارزیابی روش رونوشت برداری معکوس و واکنش تکثیری هم دما به واسطه لوپ برای شناسایی اسهال ویروسی گاو ویروس 1 و 2 | ||
| نویسندگان [English] | ||
| امیر تاجبخش1؛ الهام رضاتوفیقی1؛ خلیل میرزاده2؛ مهدی پور مهدی3 | ||
| 1گروه زیست شناسی، دانشکده علوم، دانشگاه شهید چمران اهواز، اهواز، ایران | ||
| 2گروه علوم دامی و صنایع غذایی، دانشگاه کشاورزی و منابع طبیعی رامین، اهواز، ایران | ||
| 3دانشکده دامپزشکی، دانشگاه شهید چمران اهواز، اهواز، ایران | ||
| چکیده [English] | ||
| ویروس اسهال ویروسی گاو یک عامل بیماریزا برای گاوها میباشد و دارای اهمیت اقتصادی در همه دنیا است. این عامل باعث تحمیل هزینههای فراوان به صنعت دامپروری میشود. روش تک مرحلهایی رونوشت برداری معکوس و تکثیر هم دما به واسطه لوپ (RT-LAMP) برای شناسایی سریع و موثر ویروس اسهال ویروسی گاو طراحی گردید. چهار پرایمر بر اساس ناحیه حفاظت شده غیر ترجمهایی انتهای΄5 (5΄UTR) ژنوم ویروس اسهال ویروسی گاو برای تشخیص شش محل مختلف روی RNA هدف طراحی گردید. هشتاد نمونه خون از دامهای مشکوک به اسهال ویروسی گاو جمع آوری و به طور همزمان با دو روش RT-PCR وRT-LAMP تست شدند. 24 نمونه از این تعداد با روشRT-LAMP مثبت شدند. درحالیکه با روش RT-PCR بیست نمونه مثبت شدند. حد شناسایی برای روش RT-LAMP تقریبا PFU/mL70 تخمین زده شد. مقایسه روش RT-PCR با RT-LAMP نشان داد که روش جدید طراحی شده به عنوان یک روش حساس و اختصاصی توانایی تشخیص این ویروس در نمونه های کلینیکی را دارد. | ||
| کلیدواژه‌ها [English] | ||
| اسهال ویروسی گاو, رونوشت برداری معکوس-تکثیر هم دما به واسطه لوپ, رونوشت برداری معکوس, واکنش زنجیره ایی پلیمراز | ||
| مراجع | ||
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