Detection of bacteria carrying the stx2 gene by in situ loop-mediated isothermal amplification
Detection of bacteria carrying the stx2 gene by in situ loop-mediated isothermal amplification
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DOI：
10.1128/aem.69.8.5023-5028.2003
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发表时间：
2003-08-01
影响因子：
4.4
通讯作者：
Nasu, M
中科院分区：
文献类型：
--
作者：
Maruyama, F;Kenzaka, T;Nasu, M
介绍了一种新的原位DNA扩增技术，用于显微镜下检测携带特定基因的细菌。应用环介导等温扩增（LAMP）技术检测大肠杆菌O157：H7细胞中stxA（2）。与原位PCR相比，原位LAMP方法中使用的温和的透化条件和低的等温温度引起的细胞损伤较小。在DNA扩增后的细菌混合物中，可以使用荧光抗体标记来鉴定E。coliO157：H7细胞中的stxA（2）基因。与原位PCR相比，该方法获得了更高对比度的图像。
A new in situ DNA amplification technique for microscopic detection of bacteria carrying a specific gene is described. Loop-mediated isothermal amplification (LAMP) was used to detect stxA(2) in Escherichia coli O157:H7 cells. The mild permeabilization conditions and low isothermal temperature used in the in situ LAMP method caused less cell damage than in situ PCR. It allowed use of fluorescent antibody labeling in the bacterial mixture after the DNA amplification for identification of E. coli O157:H7 cells with an stxA(2) gene. Higher-contrast images were obtained with this method than with in situ PCR.