A Novel RT-LAMP Assay for Rapid and Simple Detection of Classical Swine Fever Virus  

作　　者：Lei CHEN Xue-zheng FAN Qin WANG Lu XU Qi-zu ZHAO Yuan-chen ZHOU Jun LIU Bo TANG Xing-qi ZOU 

机构地区：[1]China Institute of Veterinary Drug Control, Beijing, 100081, China

出　　处：《Virologica Sinica》2010年第1期59-64,共6页中国病毒学（英文版）

基　　金：The National Science and Technology supporting plan of the Eleventh Five-year(2006BAD06A18 and 2006BAD06A03);Beijing Natural Science Foundation(5072041)

摘　　要：A simple and rapid assay for the detection of Classical swine fever virus(CSFV)was established using reverse transcription loop-mediated isothermal amplification(RT-LAMP).This study describes the amplification of the genomic RNA of CSFV under isothermal conditions(63℃)within one hour,using a set of six primers(two outer primers,two inner primers and two loop primers).This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR.This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV.PRRSV.SIV. PRV-PCV,thus showed a good specificity.Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition,either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye.Because RT-LAMP is low-cost and produces rapid results,it has the potential to be an excellent tool for CSFV surveillance in the field,especially in developing countries.A simple and rapid assay for the detection of Classical swine fever virus （CSFV） was established using reverse transcription loop-mediated isothermal amplification （RT-LAMP）. This study describes the amplification of the genomic RNA of CSFV under isothermal conditions （63 ℃） within one hour, using a set of six primers （two outer primers, two inner primers and two loop primers）. This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR. This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV. PRRSV. SIV PRV-PCV, thus showed a good specificity. Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition, either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye. Because RT-LAMP is low-cost and produces rapid results, it has the potential to be an excellent tool for CSFV surveillance in the field, especially in developing countries

关 键 词：Classical swine fever virus （CSFV） Reverse transcription loop-mediated isothermal amplification（RT-LAMP） Rapid detection 

分 类 号：S852.651[农业科学—基础兽医学] TP309[农业科学—兽医学]