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《Chinese Journal of Preventive Veterinary Medicine》 2009-05
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Establishment of a rapid method for detection of peste des petits ruminants virus by a reverse transcription loop-mediated isothermal amplification

LI Wei1,LI Gang1,2,FAN Xiao-juan1,ZHANG Kun1,JIA Feng-qin1,SHI Li-jun1,Hermann Unger3(1.Beijing Institute of Animal Science and Veterinary Medicine,Chinese Academy of Agricultural Sciences,Beijing 100193,China;2.State Key Laboratory of Animal Nutrition,Beijing 100094,China;China;3.FAO/IAEA Agriculture and Biotechnology Laboratory,Seibersdorf,Austria)  
A one-step reverse transcription loop-mediated isothermal amplification(RT-LAMP) assay was developed for rapid and sensitive detection of peste des petits ruminants virus(PPRV).A set of four primers were designed based on the PPRV N gene sequence;RT-LAMP was performed using Bst DNA polymerase large fragment,and ladder like DNA fragments were observed on agarose gel electrophoresis assay.The process takes only 70 minutes and the amplification results could be visualized with SYBR Green I.The sensitivity test indicated that the RT-LAMP was 1000 times higher than that of RT-PCR,while 100 times higher of nest RT-PCR.The method described in this study was proved to be a sensitive,specific and rapid way for the detection of PPRV in infected cells and tissues.It has a potential application of both laboratorial and pen-side detection of PPRV.
【Fund】: “十一五”国家科技支撑计划(2006BAD06A13-4);; 国家“863”计划(2008AA10Z411);; FAO/IAEA项目(14515-0 R1);; “948”项目(2007-G57-C)
【CateGory Index】: S854.4
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