Direct and sensitive detection of a microsporidian parasite of bumblebees using loop-mediated isothermal amplification (LAMP)
Direct and sensitive detection of a microsporidian parasite of bumblebees using loop-mediated isothermal amplification (LAMP)
复制标题
使用环介导等温扩增 (LAMP) 直接、灵敏地检测熊蜂的微孢子虫寄生虫
DOI：
10.1038/s41598-020-57909-8
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发表时间：
2020
影响因子：
4.6
通讯作者：
Inoue Maki N.
中科院分区：
文献类型：
--
作者：
Kato Yuto;Yanagisawa Takahiro;Nakai Madoka;Komatsu Ken;Inoue Maki N.
在过去的几十年里，熊蜂种群的减少已经被报道，而微孢子虫寄生虫Nosema bombis被认为是导致这种减少的因素之一。虽然蜜蜂数量的减少影响了野生植物和人类的食物供应，但微孢子虫的影响。感染是未知的，因为很难从野生蜜蜂中获得感染性孢子，因为它们的流行率很低。粪便样品或中肠匀浆的显微镜观察和/或PCR通常用于N。炸弹探测但是，如果将微孢子虫孢子长时间保存在4 °C或冷冻，其萌发率会下降。因此，至关重要的是，要尽量减少诊断和从现场采集的样本中分离感染性孢子的时间。因此，我们进行了环介导等温扩增（LAMP）检测直接检测N。熊蜂中肠匀浆。利用这种方法，我们可以检测N。从显微镜下可见的个体和直接从野生个体中分离。
The reduction of bumblebee populations has been reported in the last decades, and the microsporidian parasiteNosema bombiis considered as one of the factors contributing to such reduction. Although the decline of bee populations affects both wild plants and human food supply, the effects ofNosemaspp. infections are not known because it is difficult to obtain infective spores from wild bees due to their low prevalence. Microscopical observation of fecal samples or midgut homogenates and/or PCR are generally used forN. bombidetection. However, the germination rate of microsporidian spore declines if they are kept at 4 °C for a long time or frozen. It is therefore crucial to minimize the diagnosis and isolation time of infective spores from field-collected samples. Therefore, we performed a loop-mediated isothermal amplification (LAMP) assay for the direct detection ofN. bombiin bumblebee midgut homogenates. Using this method, we could detectN. bombifrom individuals from which it was visible under the microscope and directly from wild individuals.