کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8747030 | 1593419 | 2018 | 31 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Development of fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) using quenching probes for the detection of the Middle East respiratory syndrome coronavirus
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کلمات کلیدی
PBSHBoVMERS coronavirusHCoVTCID50ffuRT-LAMPnucleocapsidMPVRSVFIPATCCUPEAdVPIVORF50% tissue culture infectious dose - 50٪ بافت کشت عفونیBiP - BIPmERs - mers هاCOV - THEAdenovirus - آدنوویروسhuman coronavirus - انسان کرونا ویروسMiddle East respiratory syndrome - سندرم تنفسی خاورمیانهopen reading frame - قاب خواندن بازMetapneumovirus - متاپنهوم ویروسیAmerican Type Culture Collection - مجموعه فرهنگی نوع آمریکاییPhosphate-buffered saline - محلول نمک فسفات با خاصیت بافریplaque forming unit - واحد پالک تشکیل شده استHuman bocavirus - ویروس انسانی انسانRespiratory syncytial virus - ویروس سنسیتیال تنفسیParainfluenza virus - ویروس پاراآنفلوآنزاpfu - پفوCoronavirus - کروناویروس
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ویروس شناسی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Development of fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) using quenching probes for the detection of the Middle East respiratory syndrome coronavirus Development of fluorescent reverse transcription loop-mediated isothermal amplification (RT-LAMP) using quenching probes for the detection of the Middle East respiratory syndrome coronavirus](/preview/png/8747030.png)
چکیده انگلیسی
Clinical detection of Middle East respiratory syndrome (MERS) coronavirus (MERS-CoV) in patients is achieved using genetic diagnostic methods, such as real-time RT-PCR assay. Previously, we developed a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for the detection of MERS-CoV [Virol J. 2014. 11:139]. Generally, amplification of RT-LAMP is monitored by the turbidity induced by precipitation of magnesium pyrophosphate with newly synthesized DNA. However, this mechanism cannot completely exclude the possibility of unexpected reactions. Therefore, in this study, fluorescent RT-LAMP assays using quenching probes (QProbes) were developed specifically to monitor only primer-derived signals. Two primer sets (targeting nucleocapsid and ORF1a sequences) were constructed to confirm MERS cases by RT-LAMP assay only. Our data indicate that both primer sets were capable of detecting MERS-CoV RNA to the same level as existing genetic diagnostic methods, and that both were highly specific with no cross-reactivity observed with other respiratory viruses. These primer sets were highly efficient in amplifying target sequences derived from different MERS-CoV strains, including camel MERS-CoV. In addition, the detection efficacy of QProbe RT-LAMP was comparable to that of real-time RT-PCR assay using clinical specimens from patients in Saudi Arabia. Altogether, these results indicate that QProbe RT-LAMP assays described here can be used as powerful diagnostic tools for rapid detection and surveillance of MERS-CoV infections.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Virological Methods - Volume 258, August 2018, Pages 41-48
Journal: Journal of Virological Methods - Volume 258, August 2018, Pages 41-48
نویسندگان
Kazuya Shirato, Shohei Semba, Sherif A. El-Kafrawy, Ahmed M. Hassan, Ahmed M. Tolah, Ikuyo Takayama, Tsutomu Kageyama, Tsugunori Notomi, Wataru Kamitani, Shutoku Matsuyama, Esam Ibraheem Azhar,