Ohtsuki R, Kawamoto K, Kato Y, Shah M M, Ezaki T, Makino S-I
Laboratory of Food Microbiology and Immunology, Research Center for Animal Hygiene and Food Safety, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido, Japan.
J Appl Microbiol. 2008 Jun;104(6):1815-23. doi: 10.1111/j.1365-2672.2008.03732.x. Epub 2008 Jan 31.
To develop a rapid and sensitive method for detecting Brucella spp.
Two sets of six Brucella-specific primers for loop-mediated isothermal amplification (LAMP) were designed from the sequence of the Brucella abortus BCSP31 gene. The specificity and sensitivity were examined for six Brucella species (22 strains) and 18 non-Brucella species (28 strains). The LAMP assay was specific to Brucella spp. in 35 min at 63 degrees C and sensitive (detected 10 fg of genomic DNA). The assay was also applied for the detection of Brucella DNA in contaminated milk and infected mouse organs.
We developed a sensitive and specific LAMP assay for Brucella spp., with the test appearing to be useful for the detection of the pathogen from clinical and food samples.
This is the first report of the development of LAMP for the detection of Brucella spp. As the LAMP assay can be performed at a constant temperature and its reactivity is directly observed with the naked eye without electrophoresis, our assay should be useful for the diagnosis of brucellosis as well as the detection of the bacteria in environmental or food samples.
开发一种快速、灵敏的检测布鲁氏菌属的方法。
根据流产布鲁氏菌BCSP31基因序列设计了两组用于环介导等温扩增（LAMP）的六条布鲁氏菌特异性引物。对六种布鲁氏菌（22株）和18种非布鲁氏菌（28株）进行了特异性和敏感性检测。LAMP检测在63℃下35分钟对布鲁氏菌属具有特异性，且灵敏度高（可检测到10 fg基因组DNA）。该检测方法还应用于污染牛奶和感染小鼠器官中布鲁氏菌DNA的检测。
我们开发了一种针对布鲁氏菌属的灵敏且特异的LAMP检测方法，该检测方法似乎对从临床和食品样本中检测病原体有用。
这是关于开发用于检测布鲁氏菌属的LAMP的首次报道。由于LAMP检测可在恒温下进行，且无需电泳即可直接用肉眼观察其反应性，我们的检测方法应有助于布鲁氏菌病的诊断以及环境或食品样本中细菌的检测。
