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Edrisi, Mehdi, Safi, Shahabeddin, Varshovi, Hamid Reza, Shahhosseiny, Mohammad Hassan. (1403). Evaluation and optimization of loop-mediated isothermal amplification (LAMP) technique in capripoxvirus diagnosis and its comparison with PCR method. سامانه مدیریت نشریات علمی, 79(6), 1183-1190. doi: 10.32592/ARI.2024.79.6.1183
Mehdi Edrisi; Shahabeddin Safi; Hamid Reza Varshovi; Mohammad Hassan Shahhosseiny. "Evaluation and optimization of loop-mediated isothermal amplification (LAMP) technique in capripoxvirus diagnosis and its comparison with PCR method". سامانه مدیریت نشریات علمی, 79, 6, 1403, 1183-1190. doi: 10.32592/ARI.2024.79.6.1183
Edrisi, Mehdi, Safi, Shahabeddin, Varshovi, Hamid Reza, Shahhosseiny, Mohammad Hassan. (1403). 'Evaluation and optimization of loop-mediated isothermal amplification (LAMP) technique in capripoxvirus diagnosis and its comparison with PCR method', سامانه مدیریت نشریات علمی, 79(6), pp. 1183-1190. doi: 10.32592/ARI.2024.79.6.1183
Edrisi, Mehdi, Safi, Shahabeddin, Varshovi, Hamid Reza, Shahhosseiny, Mohammad Hassan. Evaluation and optimization of loop-mediated isothermal amplification (LAMP) technique in capripoxvirus diagnosis and its comparison with PCR method. سامانه مدیریت نشریات علمی, 1403; 79(6): 1183-1190. doi: 10.32592/ARI.2024.79.6.1183

Evaluation and optimization of loop-mediated isothermal amplification (LAMP) technique in capripoxvirus diagnosis and its comparison with PCR method

Archives of Razi Institute
مقاله 7، دوره 79، شماره 6، بهمن و اسفند 2024، صفحه 1183-1190    اصل مقاله (329.04 K)
نوع مقاله: Original Articles
شناسه دیجیتال (DOI): 10.32592/ARI.2024.79.6.1183
نویسندگان
Mehdi Edrisi1؛ Shahabeddin Safi* 1؛ Hamid Reza Varshovi2؛ Mohammad Hassan Shahhosseiny3
1Department of Pathobiology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
3Department of Microbiology, Shahr-e-Qods Branch, Islamic Azad University, Tehran, Iran
چکیده
Sheep pox (SP), goat pox (GP), and lumpy skin disease (LSD), subspecies of the Capripox virus (CaPVs), are important in the pathogenesis of sheep, goats, and cattle. The causative agent is the capripox virus which has been isolated in South Africa for the first time. Sheeppox (SP), goatpox (GP), and lumpy skin disease (LSD) viruses are morphologically indistinguishable from each other and have been adapted to different host species (4). From a serological point of view, it is difficult to differentiate these viruses and cross-immunity occurs among them (2). The present study reports the evaluation and optimization of a novel loop-mediated isothermal amplification (LAMP) technique for the rapid detection of CaPVs and compares the LAMP technique with the PCR method. LAMP primers were selected from the P32-protected gene of the Capripox virus. Safe-Red fluorescent dye was used to monitor and change the color in the disease’s positive cases to bright yellow at a 320-nm wavelength, and the final results were confirmed using electrophoresis. The proposed LAMP test for the Capripox virus showed high specificity and non-cross-reactivity with viruses of Poxviridae with similar clinical symptoms. The optimized LAMP test was compared with PCR. LAMP and PCR were similar in diagnostic sensitivity. The specificity was evaluated using 30 samples of cow skin suspected of having lumpy skin disease as well as 16 samples of positive and negative references and the negative control. The proposed easy-to-use, inexpensive, high-sensitivity LAMP test might be suitable for detecting caprypox virus in laboratories in border areas and rural areas.
کلیدواژه‌ها
LSDV؛ LAMP؛ PCR؛ Capripoxvirus
عنوان مقاله [English]
ارزیابی و بهینه سازی تکنیک Loop-mediated isothermal amplification (LAMP) درتشخیص capripoxvirus و مقایسه آن با روش PCR
چکیده [English]
آبله گوسفند (SP) ، آبله بز(GP) و بیماری لمپی اسکین (LSD) از زیر گونه capripoxvirus (CaPVs) می باشند که اهمیت اقتصادی در بیماری زایی گوسفند ، بز و گاو برعهده دارند. ما در اینجا ارزیابی و بهینه سازی تکنیک بی همتای LAMP را برای تشخیص سریع کاپری پاکس ویروس و مقایسه آن با روش PCR را گزارش می کنیم. پرایمرهای LAMP از ژنوم محافظت شده P32 از این جنس انتخاب شدند. رنگ فلوروسنتی SYBR safe برای مانیتورینگ و ایجاد تغییر رنگ در موارد مثبت بیماری به سبز درخشان در طول موج 350 نانومتر بکار گرفته شد و نتایج نهایی با الکتروفورزیس تأیید شد. تست LAMP با ویژگی بالا برای capripoxvirus و عدم واکنش متقاطع حتی با ویروس های خانواده پاکس ویریده و یا ویروس های با علایم کلینیکی مشابه ارزیابی شد. تست LAMP بهینه شده با روش PCR مقایسه شد. LAMP و PCR از لحاظ حساسیت تشخیصی مشابه و با محدوده تشخیص LOD 3 و 8 کپی ژنوم می باشد. ویژگی تشخیصی با تعداد 30 نمونه پوست گاو مشکوک به بیماری لمپی اسکین و 16 عدد نمونه رفرانس مثبت و منفی و کنترل منفی ارزیابی شد . تست LAMP با قابلیت استفاده آسان ، ارزان با حساسیت بالا و بخصوص مناسب برای تشخیص کاپری پاکس ویروس در آزمایشکاه های مناطق مرزی و آزمایشگاه های مناطق روستایی، در این گزارش توصیف می شود.
کلیدواژه‌ها [English]
ویروس لمپی اسکین, LAMP, واکنش زنجیره ای پلمیراز, کاپری پاکس ویروس
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