中国水稻科学2017，Vol.31Issue(4)：432-440,9.DOI:10.16819/j.1001-7216.2017.7001
水稻干尖线虫的环介导恒温扩增技术(LAMP)快速检测方法
Loop-mediated Isothermal Amplification Assay for Rapid Diagnosis of Aphelenchoides besseyi
摘要
Abstract
[Objective]This study was performed to provide a new method for the rapid detection and early diagnosis of Aphelenchoides besseyi. [Method]The loop-mediated isothermal amplification (LAMP) specific primers of A. besseyi were designed based on the nematode 18S ribosomal RNA gene, including two external primer, two inner primer and two loop primer. The DNA sample was amplified by LAMP primers using isothermal amplification. The amplified products were detected by electrophoresis and fluorescent dye. It proved that there was A. besseyi in the test sample if there was clear DNA ladder in the electrophoresis results or it showed green fluorescence in the fluorescent dye results. [Result]The results showed that LAMP primers could detect and identify single egg, juvenile, female and male of A. besseyi, and could also directly detect A. besseyi from multiple nematode species samples and plant tissue samples, the sensitivity of LAMP assay was 1/1000 of single nematode DNA. [Conclusion]This diagnosis method has the advantages of accuracy, sensitivity, stability and intuitive results, and is also simple and more practical.关键词
水稻干尖线虫/LAMP/快速检测/灵敏度Key words
Aphelenchoides besseyi/LAMP/rapid diagnosis/sensitivity分类
农业科技引用本文复制引用
白宗师,秦萌,赵立荣,韩玉春,王东伟,徐春玲,谢辉..水稻干尖线虫的环介导恒温扩增技术(LAMP)快速检测方法[J].中国水稻科学,2017,31(4):432-440,9.基金项目
国家自然科学基金资助项目(31371920). （31371920）
