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《Progress in Veterinary Medicine》 2017-12
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Development of a Novel RT-LAMP Assay for Detection of African Horse Sickness Virus

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;  
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.
【Fund】: “十二五”国家科技支撑计划项目(2013BADl2804)
【CateGory Index】: S852.652
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