Melissococcus plutonius is the causative agent of European foulbrood of honey bee larvae (Apis mellifera). The early diagnosis of disease is necessary for its elimination. Previously, we found false-positive results for PCR detection of M. plutonius in honey bee workers by qPCR. Here we report the comparison of PCR methods for quantification of M. plutonius in the bacteria spiked samples of honey bee workers and detection limits to eliminate false-positive results. We compare available primers of qPCR and PCR detection. The obtained detection limit was 10² copies of M. plutonius. The results of this comparison show conventional PCR detection using EFB_primers as a suitable method to confirm visual observation of European foulbrood using samples of honey bee workers. Based on our estimation, the numbers of M. plutonius should be quantified by copies of the Naþ/H þ 50 antiporter marker (MP_primers) in honey bee worker samples

Melissococcus plutonius是欧洲蜜蜂幼虫（Apis mellifera）的臭虫的病原体。疾病的早​​期诊断对于消除它是必要的。以前，我们发现通过qPCR PCR检测蜜蜂工人中的M. plutonius时出现假阳性结果。在这里，我们报告了定量PCR方法的比较，该方法定量了蜜蜂工蜂的加标细菌样品中的M. plutonius和检测限，以消除假阳性结果。我们比较了qPCR和PCR检测的可用引物。所获得的检出限为10 ^2个拷贝的M. plutonius。比较结果表明使用EFB_primers进行常规PCR检测作为确认使用蜜蜂工人样品对欧洲钩针动物进行肉眼观察的合适方法。根据我们的估计，应通过蜜蜂工人样本中Naþ/ Hþ50反转运蛋白标记（MP_primers）的副本来定量检测plutonius的数量。