Development of a fast and reliable diagnostic method for American foulbrood disease (Paenibacillus larvae subsp larvae) using a 16S rRNA gene based PCR
Development of a fast and reliable diagnostic method for American foulbrood disease (Paenibacillus larvae subsp larvae) using a 16S rRNA gene based PCR
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DOI：
10.1051/apido:2001136
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发表时间：
2001-07-01
期刊：
影响因子：
2.4
通讯作者：
Jacobs, FJ
中科院分区：
文献类型：
--
作者：
Dobbelaere, W;de Graaf, DC;Jacobs, FJ
幼虫类芽孢杆菌（Paenibacilluslarvae）是蜜蜂的一种严重疾病，是引起蜜蜂美洲幼虫病（AmericanFoulbrood，AFB）的病原.及时检测和识别这种微生物的能力对于在AFB情况下采取有效的卫生措施非常重要。P. larvae幼虫是一种难养且生长缓慢的细菌，初步分离和推定鉴定可能困难且耗时。在这项研究中，使用PCR描述了一个快速和可靠的诊断。所开发的PCR检测方法对疟原虫幼虫具有特异性，因为13种相关或蜂箱相关物种均未产生扩增子。仅以P.larvae幼虫和P.larvae pulvifaciens DNA为模板，形成扩增子。直接从AFB病幼虫尸体中提取DNA也观察到阳性反应。因此，该PCR检测仅在4小时内就提供了可靠的AFB诊断。
Paenibacillus larvae larvae is the causative agent of American foulbrood (AFB), a serious honeybee disease. The ability to detect and identify this organism promptly is important to allow effective sanitary measures in case of AFB. P. larvae larvae is a fastidious and slow-growing bacterium and primary isolation and presumptive identification can be difficult and time-consuming. In this study the use of PCR is described for a rapid and reliable diagnosis. The developed PCR assay is specific for P. larvae as no amplicons were produced from 13 related or hive-related species. Only with P. larvae larvae and P. larvae pulvifaciens DNA as template, an amplicon was formed. A positive reaction was also observed when DNA was extracted directly from remains of an AFB diseased larva. Thus this PCR assay provides a reliable diagnosis for AFB in only 4 hours.