A PCR-based method that permits specific detection of Paenibacillus larvae subsp larvae, the cause of American Foulbrood of honey bees, at the subspecies level
A PCR-based method that permits specific detection of Paenibacillus larvae subsp larvae, the cause of American Foulbrood of honey bees, at the subspecies level
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DOI：
10.1111/j.1472-765x.2004.01535.x
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发表时间：
2004-01-01
影响因子：
2.4
通讯作者：
Aguilar, OM
中科院分区：
文献类型：
--
作者：
Alippi, AM;López, AC;Aguilar, OM
目的：建立一种可靠的芽孢杆菌幼虫亚种鉴定方法。蜜蜂(Apis melvena L.)Foulbrood病的病原幼虫。方法和结果：采用ERIC-PCR方法，在不同菌株间扩增出约970bp的扩增片段，而在其他亲缘关系较近的物种中未发现扩增片段。根据该扩增片段的核苷酸序列数据，我们设计了一对寡核苷酸Kat 1和Kat 2，并将其作为引物进行了聚合酶链式反应。结论：该方法可对L拟青霉幼虫进行特异性检测。本研究的意义和影响：该方法能区分拟青霉幼虫亚种，具有高度的特异性。近缘拟青霉幼虫亚种的幼虫。普罗维菲斯。该技术可以直接用于检测蜜蜂孵化样品和受污染的蜂蜜中是否存在白纹伊蚊幼虫孢子。
Aims: A reliable procedure for the identification of Paenibacillus larvae subsp. larvae, the causal agent of American Foulbrood disease of honey bees (Apis mellifera L.) based on the polymerase chain reaction (PCR) and subspecies - specific primers is described.Methods and Results: By using ERIC-PCR, an amplicon of ca 970 bp was found among P. l. larvae strains but not in other closely related species. Based on the nucleotide sequence data of this amplicon, we designed the pair of oligonucleotides KAT 1 and KAT 2, which were assayed as primers in a PCR reaction. A PCR amplicon of the expected size ca 550 bp was only found in P. l. larvae strains.Conclusions: This PCR assay provides a specific detection for P. l. larvae.Significance and Impact of the Study: The developed PCR assay is highly specific because can differentiate Paenibacillus larvae subsp. larvae from the closely related Paenibacillus larvae subsp. pulvifaciens. The technique can be directly used to detect presence or absence of P. l. larvae spores in honey bee brood samples and contaminated honeys.