您好，欢迎访问福建省农业科学院 机构知识库！
福建省农业科学院机构知识库

全部

  • 全部
  • 标题
  • 学者
  • 机构
  • 关键词

Comparative Evaluation of the LAMP Assay and PCR-Based Assays for the Rapid Detection of Altemaria solani

文献类型： 外文期刊

作者： Khan, Mehran 1 ; Wang, Rongbo 1 ; Li, Benjin 1 ; Liu, Peiqing 1 ; Weng, Qiyong 1 ; Chen, Qinghe 1 ;

作者机构： 1.Fujian Acad Agr Sci, Fujian Key Lab Monitoring & Integrated Management, Inst Plant Protect, Fuzhou, Fujian, Peoples R China

2.Fujian Agr & Forestry Univ, State Key Lab Ecol Pest Control Fujian & Taiwan C, Fuzhou, Fujian, Peoples R China

3.Fujian Agr & Forestry Univ, State Key Lab Ecol Pes

关键词： Altemaria solani; early blight; histidine kinase gene; LAMP; real-time PCR; sensitivity; specificity

期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：5.64； 五年影响因子：6.32 ）

ISSN： 1664-302X

年卷期： 2018 年 9 卷

页码：

收录情况： SCI

摘要： Early blight (EB), caused by the pathogen Altemaria solani, is a major threat to global potato and tomato production. Early and accurate diagnosis of this disease is therefore important. In this study, we conducted a loop-mediated isothermal amplification (LAMP) assay, as well as conventional polymerase chain reaction (PCR), nested PCR, and quantitative real-time PCR (RT-qPCR) assays to determine which of these techniques was less time consuming, more sensitive, and more accurate. We based our assays on sequence-characterized amplified regions of the histidine kinase gene with an accession number (FJ424058). The LAMP assay provided more rapid and accurate results, amplifying the target pathogen in less than 60 min at 63 degrees C, with 10-fold greater sensitivity than conventional PCR. Nested PCR was 100-fold more sensitive than the LAMP assay and 1000-fold more sensitive than conventional PCR. qPCR was the most sensitive among the assays evaluated, being 10-fold more sensitive than nested PCR for the least detectable genomic DNA concentration (100 fg). The LAMP assay was more sensitive than conventional PCR, but less sensitive than nested PCR and qPCR; however, it was simpler and faster than the other assays evaluated. Despite of the sensitivity, LAMP assay provided higher specificity than qPCR. The LAMP assay amplified A. solani artificially, allowing us to detect naturally infect young potato leaves, which produced early symptoms of EB. The LAMP assay also achieved positive amplification using diluted pure A. solani culture instead of genomic DNA. Hence, this technique has greater potential for developing quick and sensitive visual detection methods than do other conventional PCR strategies for detecting A. solani in infected plants and culture, permitting early prediction of disease and reducing the risk of epidemics.

  • 相关文献
作者其他论文 更多>>
置顶
购物车
反馈

© 京ICP备09089781号-29 主办单位：福建省农业科学院

学术资源： 中国农业科学院 农业专业知识服务系统 农科联盟资源共建共享平台 中国农业科技文献与信息服务平台 中国农业期刊集成服务平台