作 者：
卢冰霞;刘磊;秦毅斌;赵武;梁家幸;郭旋;黎木兰;李斌;何颖
单 位：
广西大学动物科学技术学院;广西兽医研究所
关键词：
猪流行性乙型脑炎病毒;RT-LAMP;敏感性;特异性
摘 要：
[目的]建立1种快速、灵敏、特异的检测猪流行性乙型脑炎病毒(JEV)逆转录环介导等温扩增(RT-LAMP)检测方法。[方法]根据JEV E基因序列的保守区6个位点设计了4条特异性LAMP引物,以JEV阳性样品RNA为模板进行一步法扩增,并对反应条件和反应体系进行了优化。[结果]该方法具有较高的灵敏度,其检测极限为0.5 pg;特异性试验表明其具有较高的特异性,对其他病毒性病原体均无扩增反应。与RT-Nested-PCR相比,2种方法检出率的符合率为90.9%。反应结束后可以通过添加化学发光剂进行可视化观察,大大缩短了检测时间。该方法无需特殊仪器,只需在水浴锅中进行,是一种适于基层的简便、灵敏、快速的猪JE的检测方法。[结论]成功建立了猪流行性乙型脑炎RT-LAMP方法,并在临床上进行了初步应用。
译 名：
The Construction and Application of the Method of RT-LAMP for Swine Japanese Encephalitis
作 者：
LU Bing-xia et al(Guangxi Veterinary Research Institute,Nanning,Guangxi 530001)
关键词：
Swine Japanese B encephalitis;RT-LAMP;Sensitivity;Specificity
摘 要：
[Objective] To establish a rapid,sensitive and specific detection method which is reverse transcription loop-mediated isothermal amplification(RT-LAMP) detection method for porcine Japanese encephalitis virus(JEV).[Method] Four specificity LAMP primers were designed according to the conserved region of the six loci of the JEV E gene sequence,JEV positive sample RNA as a template for one-step amplification,and the reaction conditions and the reaction system was optimized.[Result]The results showed that the method had high sensitivity,the detection limit was 0.5 pg;specificity experiments proved that with high specificity and there was no amplification reactions for other viral pathogens.Compared with the RT-Nested-PCR,the detection rate was 90.9%.And by adding a chemiluminescent agent after the end of the reaction,could conduct visual observation,greatly reducing the detection time.This method did not require special equipment,just in the water bath,it will be a sensitive and rapid detection method for swine JE,that should be a simple suitable for grassroots.[Conclusion] A swine Japanese encephalitis RT-LAMP method was successfully established,and preliminarily used in clinical.
