Agricultural College, Guangdong Ocean University, Zhanjiang, China.
Curr Microbiol. 2011 May;62(5):1400-4. doi: 10.1007/s00284-011-9872-x. Epub 2011 Jan 28.
A set of six specific primers was designed by targeting intergenic spacer region (IGS) sequences. With Bst DNA polymerase, the products could be clearly amplified for 60 min at 62 °C in a simple water bath. The sensitivity of the loop-mediated isothermal amplification (LAMP) for detecting Metarhizium anisopliae var. anisopliae was about 0.01 pg fungal DNA per reaction (equivalent to 27 conidia). LAMP products could be judged with agar gel or naked eye after addition of SYBR Green I. There were no cross reactions with other fungal isolates indicating high specificity of the LAMP. The LAMP could detect the presence of M. anisopliae var. anisopliae from soil. The detection limits for M. anisopliae var. anisopliae of LAMP reaction was 50 conidia per reaction in soil.
设计了一套六对针对种间间隔区（IGS）序列的特异性引物。在 Bst DNA 聚合酶的作用下，在简单的水浴中于 62°C 下 60 分钟即可清晰扩增产物。检测金龟子绿僵菌 var. anisopliae 的环介导等温扩增（LAMP）的灵敏度约为每个反应 0.01pg 真菌 DNA（相当于 27 个分生孢子）。加入 SYBR Green I 后，可以用琼脂凝胶或肉眼判断 LAMP 产物。与其他真菌分离物无交叉反应，表明 LAMP 具有高度特异性。LAMP 可从土壤中检测到金龟子绿僵菌 var. anisopliae 的存在。LAMP 反应检测土壤中金龟子绿僵菌 var. anisopliae 的检测限为每个反应 50 个分生孢子。
