一种用于快速鉴定蜜蜂幼虫中蜂房蜜蜂球菌的聚合酶链式反应（PCR）检测方法。
A PCR detection method for rapid identification of Melissococcus pluton in honeybee larvae.
作者信息
Govan V A, Brözel V, Allsopp M H, Davison S
机构信息
Department of Microbiology, University of the Western Cape, Bellville, South Africa.
出版信息
Appl Environ Microbiol. 1998 May;64(5):1983-5. doi: 10.1128/AEM.64.5.1983-1985.1998.
Melissococcus pluton is the causative agent of European foulbrood, a disease of honeybee larvae. This bacterium is particularly difficult to isolate because of its stringent growth requirements and competition from other bacteria. PCR was used selectively to amplify specific rRNA gene sequences of M. pluton from pure culture, from crude cell lysates, and directly from infected bee larvae. The PCR primers were designed from M. pluton 16S rRNA sequence data. The PCR products were visualized by agarose gel electrophoresis and confirmed as originating from M. pluton by sequencing in both directions. Detection was highly specific, and the probes did not hybridize with DNA from other bacterial species tested. This method enabled the rapid and specific detection and identification of M. pluton from pure cultures and infected bee larvae.
蜂房蜜蜂球菌是欧洲幼虫腐臭病的病原体，这是一种蜜蜂幼虫的疾病。由于其严格的生长要求以及来自其他细菌的竞争，这种细菌特别难以分离。采用聚合酶链反应（PCR）从纯培养物、粗细胞裂解物以及直接从受感染的蜜蜂幼虫中选择性地扩增蜂房蜜蜂球菌的特定核糖体RNA（rRNA）基因序列。PCR引物是根据蜂房蜜蜂球菌16S rRNA序列数据设计的。PCR产物通过琼脂糖凝胶电泳进行可视化，并通过双向测序确认为源自蜂房蜜蜂球菌。检测具有高度特异性，探针不与所测试的其他细菌物种的DNA杂交。该方法能够从纯培养物和受感染的蜜蜂幼虫中快速、特异性地检测和鉴定蜂房蜜蜂球菌。
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