Department of Entomology, Michigan State University, 4090 N. College Road, Pollinator Performance Center, RM 100, Lansing, MI 48910, USA.
Department of Comparative Medicine and Integrative Biology, Veterinary Medical Center, Michigan State University, 784 Wilson Road, Room G100, East Lansing, MI 48824, USA.
J Insect Sci. 2021 Nov 1;21(6). doi: 10.1093/jisesa/ieab075.
One of the most serious bacterial pathogens of Western honey bees (Apis mellifera Linnaeus [Hymenoptera: Apidae]) is Melissococcus plutonius, the cause of the disease European foulbrood. Because European foulbrood is highly variable, with diverse outcomes at both the individual and colony levels, it is difficult to diagnose through visual inspection alone. Common lab diagnostic techniques include microscopic examination and molecular detection through PCR. In 2009, a lateral flow device was developed and validated for field diagnosis of European foulbrood. At the time, M. plutonius was thought to be genetically homogenous, but we have subsequently learned that this bacterium exists as multiple strains, including some strains that are classified as 'atypical' for which the lateral flow device is potentially less effective. These devices are increasingly used in the United States, though they have never been validated using strains from North America. It is essential to validate this device in multiple locations as different strains of M. plutonius circulate in different geographical regions. In this study, we validate the field use of the lateral flow device compared to microscopic examination and qPCR on larval samples from 78 commercial honey bee colonies in the United States with visual signs of infection. In this study, microscopic diagnosis was more sensitive than the lateral flow device (sensitivity = 97.40% and 89.47%, respectively), and we found no false positive results with the lateral flow device. We find high concurrence between the three diagnostic techniques, and all three methods are highly sensitive for diagnosing European foulbrood.
西方蜜蜂(Apis mellifera Linnaeus [膜翅目:蜜蜂科])最严重的细菌性病原体之一是乳球菌 plutonius,它是欧洲幼虫腐臭病的病原体。由于欧洲幼虫腐臭病变化多样,个体和群体水平的结果各不相同,仅凭肉眼观察很难诊断。常见的实验室诊断技术包括显微镜检查和通过 PCR 进行分子检测。2009 年,开发并验证了一种用于现场诊断欧洲幼虫腐臭病的横向流动装置。当时,人们认为乳球菌 plutonius 在遗传上是同质的,但我们后来了解到,这种细菌存在多种菌株,包括一些被归类为“非典型”的菌株,而横向流动装置对这些菌株的检测效果可能较差。这些装置在美国越来越多地使用,但从未使用过来自北美的菌株进行验证。在不同地点验证该设备至关重要,因为不同的乳球菌 plutonius 菌株在不同的地理区域传播。在这项研究中,我们验证了横向流动装置在有肉眼可见感染迹象的 78 个美国商业蜂群的幼虫样本上与显微镜检查和 qPCR 相比的现场使用情况。在这项研究中,显微镜诊断比横向流动装置更敏感(敏感性分别为 97.40%和 89.47%),并且我们没有发现横向流动装置的假阳性结果。我们发现这三种诊断技术具有高度一致性,并且这三种方法都非常敏感,可用于诊断欧洲幼虫腐臭病。