Rapid and reliable detection of phytoplasma by loop-mediated isothermal amplification targeting a housekeeping gene.
Rapid and reliable detection of phytoplasma by loop-mediated isothermal amplification targeting a housekeeping gene.
复制标题
通过针对管家基因的环介导等温扩增快速可靠地检测植原体。
DOI：
10.1007/s10327-012-0403-9
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发表时间：
2012
影响因子：
1.2
通讯作者：
Oshima K Namba S
中科院分区：
文献类型：
--
作者：
Sugawara K. ;Himeno M.;Keima T.;Kitazawa Y. ;Maejima K.;Oshima K Namba S
Phytoplasmas are plant pathogenic bacteria that infect more than 700 plant species. Because phytoplasma-resistant cultivars are not available for the vast majority of crops, the most common practice to prevent phytoplasma diseases is to remove infected plants. Therefore, developing a rapid, accurate diagnostic method to detect a phytoplasma infection is important. Here, we developed a phytoplasma detection assay based on loop-mediated isothermal amplification (LAMP) by targeting thegroELgene and 16S rDNA. We designed 19 primer sets for the LAMP assay and evaluated their amplification efficiency, sensitivity, and spectra to select the most suitable primer sets to detectCandidatusPhytoplasma asteris. As a result, DNA was efficiently amplified by one of the primer sets targeting thegroELgene, and LAMP assay sensitivity with this primer set was 10-fold higher than that of the polymerase chain reaction. Moreover, thegroELgene was successfully amplified from several strains ofCa.Phytoplasma asteris by this primer set, indicating that thegroELgene can be used as a LAMP assay target gene for a broad range of phytoplasma strains. Additionally, a simple DNA extraction method that omits the homogenizing and phenol extraction steps was combined with the LAMP assay to develop a simple, rapid, and convenient diagnostic method for detecting phytoplasma.
