Neoparamoeba perurans is the causative agent of amoebic gill disease (AGD). Two loop‐mediated isothermal amplification (LAMP) assays targeting the parasite 18S rRNA and the Atlantic salmon EF1α, used as internal control, were designed. The N. perurans LAMP assay did not amplify close relatives N. pemaquidensis and N. branchiphila , or the host DNA. This assay detected 10⁶ copies of the parasite 18S rRNA gene under 13 min and 10³ copies under 35 min. Five “fast‐and‐dirty” DNA extraction methods were compared with a reference method and further validated by TaqMan™ qPCR. Of those, the QuickExtract buffer was selected for field tests. Seventy‐one non‐lethal gill swabs were analysed from AGD‐clinically infected Atlantic salmon. The pathogen was detected under 23 min in fish of gill score >2 and under 39 min for lower gill scores. About 1.6% of the tests were invalid (no amplification of the internal control). 100% of positives were obtained from swabs taken from fish showing gill score ˃3, but only ~50% of positives for lower gill scores. The present LAMP assay could be implemented as a point‐of‐care test for the on‐site identification of N. perurans ; however, further work is required to improve its performance for lower scores.

中文翻译：

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非致死性环介导的等温扩增测定法可作为阿米巴bic虫病的病原体新百日草的即时诊断工具。

百日草新巴莫巴是阿米巴g病（AGD）的病原体。设计了两个针对寄生虫18S rRNA和大西洋鲑EF1α的环介导的等温扩增（LAMP）分析，用作内部对照。该N. perurans LAMP检测没有扩增近亲N. pemaquidensis和N. branchiphila，或宿主DNA。该检测在13分钟和10 ³下检测到10 ⁶个寄生虫18S rRNA基因拷贝在35分钟内复制。将五种“快速和肮脏”的DNA提取方法与参考方法进行了比较，并通过TaqMan™qPCR进行了进一步验证。其中，选择了QuickExtract缓冲区进行现场测试。从AGD临床感染的大西洋鲑鱼中分析了71个非致命性g拭子。在23评分> 2的鱼中，在23分钟以下检测到病原体，在g评分较低的鱼中，在39分钟以下检测到病原体。约1.6％的测试无效（内部对照无扩增）。100％的阳性结果是从鱼fish中抽出的，其g得分为˃3，而较低的g得分只有〜50％的阳性。目前的LAMP检测可作为现场鉴定猪毛猪笼草的即时检验。但是，需要做进一步的工作以提高其性能以降低分数。