东北林蛙种蛙蛙病毒LAMP快速检测

Rapid Detection Method of Ranaviruses from Breeding Rana dybowskii Using Loop-mediated Isothermal Amplification

由于环境、气候的恶化以及疾病的侵害,全球两栖动物种群数量呈现出整体衰退的趋势,其中虹彩病毒属蛙病毒科的蛙病毒是不可忽略的原因之一.环介导等温扩增技术（loop-mediated-isothermal-amplification,LAMP）是一种新型的核酸扩增技术,具有特异性强、快速、简便等特点,本研究利用LAMP技术,开发了针对中国东北林蛙种蛙蛙病毒的快速检测方法,针对蛙病毒MCP基因设计出能特异识别靶序列上6个位点的4条特异性引物,在链置换聚合酶（Bst酶）的作用下,62℃扩增30 min.通过对实验参数的优化,扩增后病毒的最低检出数可达到102个拷贝.该方法可用于东北林蛙养殖厂种蛙蛙病毒的临床快速检测,保证种蛙的存活质量和数量.

Due to deteriorating environmental conditions, climate change, and disease, populations of amphibians have shown global declines. Ranaviruses of the Iridoviridea are pathogens that cannot be ignored. Loop-mediated isothermal amplification （LAMP）is a new nucleic acid amplification technology, which is highly specific, rapid and simple. This study developed a rapid detection method using LAMP technology to identify ranavirus from breeding Rana dybowskii. We designed four specific primer sets to identify six fragments of the ranavirus MCP sequence. The reaction of LAMP was completed within 30 min by Bst polymerase catalysis under isothermal condition at 62℃. By optimizing the experimental parameters, the number of the minimum detectable amplification can be up to 100 copies of ranavirus. This method can be used for laboratory and clinical detection of ranavirus.