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

查看全文 作  者:Lei CHEN Xue-zheng FAN Qin WANG Lu XU Qi-zu ZHAO Yuan-chen ZHOU Jun LIU Bo TANG Xing-qi [1]ZOU 高影响力作者 机构地区:[1]China Institute of Veterinary Drug Control, Beijing, 100081, China高影响力机构 出  处:《Virologica Sinica》索引2010年第25卷第1期,共6页高影响力期刊 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. 关 键 词:猪瘟病毒 病毒检测 逆转录 琼脂糖凝胶电泳 牛病毒性腹泻 循环使用 投资工具
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