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当前位置: 首页 > 文章 > 非洲马瘟病毒RT-LAMP检测方法的建立 动物医学进展 2017 (12) 1-5
Position: Home > Articles > Development of a Novel RT-LAMP Assay for Detection of African Horse Sickness Virus Progress in Veterinary Medicine 2017 (12) 1-5

非洲马瘟病毒RT-LAMP检测方法的建立

作  者：
姜睿姣;邬旭龙;张鹏飞;王印;杨泽晓;姚学萍
关键词：
非洲马瘟病毒;环介导等温扩增;检测
摘  要：
为了建立非洲马瘟病毒(AHSV)环介导等温扩增(loop-mediated isothermal amplification,LAMP)快速检测方法,基于AHSV VP7基因保守序列,设计2对特异性扩增引物。通过对反应体系中各组分浓度,反应温度及时间的优化,建立了快速、灵敏的AHSV RT-LAMP检测方法,并对该方法特异性、灵敏度、重复性进行探索。结果表明,在63℃恒温下反应45min,便可进行高效率的特异性扩增。反应产物经琼脂糖凝胶电泳和染料可视化鉴定,能够快速有效检测AHSV。用建立的方法检测马属动物易感的4种疫病病原,结果均为阴性,证实具有较高的特异性。灵敏度是RT-PCR的1 000倍。建立了AHSV RT-LAMP检测方法,具有快速、特异、灵敏、操作简单、设备要求低的特点,适合用于现场AHSV快速检测。
译  名：
Development of a Novel RT-LAMP Assay for Detection of African Horse Sickness Virus
作  者：
JIANG Rui-jiao;WU Xu-long;ZHANG Peng-fei;WANG Yin;YANG Ze-xiao;YAO Xue-ping;College of Veterinary Medicine,Sichuan Agricultural University;Key Laboratory of Animal Disease and Human Health of Sichuan Province;
单  位：
College of Veterinary Medicine,Sichuan Agricultural University%Key Laboratory of Animal Disease and Human Health of Sichuan Province
关键词：
African horse sickness virus;;RT-LAMP;;detection
摘  要：
The objective of the study was to establish a rapid,sensitive one-step reverse transcription loopmediated isothermal amplification(RT-LAMP)assay for detecting African horse sickness virus(AHSV).Based on the AHSV VP7 gene conserved sequence,two pairs of optimal primers were designed.Through optimizing temperature,time and all components concentration in the reaction system,a rapid and specific detection of AHSV was established.The results showed that the method of LAMP manifested a highly efficient amplification for AHSV viral target gene when the reaction system was placed in 63℃ for 45 min.Reaction products could be quickly and effectively detected by agarose gel electrophoresis and dye visual identification.The established method was used to detect other susceptible diseases,eventually the results were negative.It confirmed that the method has high specificity.What's more,the detection limit was 10 copies/μL,which was approximately a 10-fold greater sensitivity than RT-PCR.Therefore,a specific,sensitive and rapid RT-LAMP detection was established,which is suitable for rapid detection with simple conditions.

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