Genome-guided, field-deployable loop-mediated isothermal amplification (LAMP) assay for specific detection ofDickeya dadantii

Author:

Montesinos Stefania,Tyagi Garima,Feng Zhuokun,Hampson Ella,Adhikari Achyut,Minaai Michael,Wong Landon,Haubner Meagan,Dobhal ShefaliORCID,Arizala Dario,Andreason Sharon A.,Mollov Dimitre,Ochoa-Corona FranciscoORCID,Bingham Jon-Paul,Odani Jenee,Jenkins Daniel,Ma Li Maria,Fletcher Jacqueline,Stack James P.,Arif MohammadORCID

Abstract

ABSTRACTPotatoes, among the most economically significant crops worldwide, are susceptible to various plant pathogens that significantly impact their propagation, production, storage, and distribution. Soft rot disease, caused primarily byDickeyaandPectobacterium, results in substantial economic losses to the agricultural industry annually. In this study, we developed a rapid, reliable, and field-deployable loop-mediated isothermal amplification (LAMP) assay for detectingD. dadantii, a common soft rot causing bacteria. TheD. dadantii-specific LAMP primers were designed targeting a highly conserved genomic region withinD. dadantii, the TetR/AcrR family transcriptional regulator CDS and its flanking sequences. This assay was thoroughly validated with the members of inclusivity (nine strains ofD. dadantii) and exclusivity panels (85 strains, including allDickeyaspecies, related taxa, and host DNA), detecting no false positives or negatives. The limit of detection (LOD) was established by performing assays with 10-fold serially diluted pure gDNA ofD. dadantiiand gDNA spiked with host crude extract; the assay detected the target pathogen down to 1 pg (188 copies) without being adversely affected by the host crude extract. The developed LAMP assay specifically detected the target pathogen in infected plant materials. Additional multi-operator blind and multi-instrument tests were conducted to assess the assay’s robustness and applicability, consistently yielding accurate results without false positives or negatives. These findings demonstrate the assay’s potential utility for biosecurity, routine diagnostics, and epidemiological studies.

Publisher

Cold Spring Harbor Laboratory

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3