Genome-Informed Loop-Mediated Isothermal Amplification (LAMP) Assays for Detection and Differentiation of Ralstonia pseudosolanacearum Phylotypes I and III
- 1Department of Entomology and Plant Pathology, North Carolina State University, Raleigh, NC 27607, USA
- 2Plant Pathogen Confirmatory Diagnostics Laboratory (PPCDL), Science and Technology (S&T), Plant Protection and Quarantine (PPQ), Animal and Plant Health Inspection Service (APHIS), United States Department of Agriculture (USDA), Laurel, MD 20708, USA
- 3Floral and Nursery Plants Research Unit, United States National Arboretum, USDA Agricultural Research Service (ARS), Beltsville, MD 20705, USA
Abstract
The Ralstonia solanacearum species complex (RSSC) causes bacterial wilt in a broad range of economically important hosts. The RSSC has been reclassified into three species, of which R. solanacearum sequevars 1 and 2 in phylotype IIB are designated as Select Agents in the U.S. Many rapid tests for “R. solanacearum” target the RSSC and thus cross-react with R. pseudosolanacearum, which is endemic to the U.S. To avoid false positive identifications and unnecessary triggering of Select Agent measures, four portable LAMP-based screening assay variations were developed and validated for specific detection of R. pseudosolanacearum: real-time LAMP assays for detection of R. pseudosolanacearum (1) and differentiation of R. pseudosolanacearum phylotypes I and III in the AmpliFire® Isothermal Fluorometer (2) or Genie® III (3), and a colorimetric LAMP for detection of R. pseudosolanacearum (4). All assays demonstrated high sensitivity (10–50 colony-forming units (CFU)/reaction) and specificity; 100% analytical specificity and selectivity based on 177 RSSC isolates (81 phylotype I, 70 phylotype II, 15 phylotype III and 11 phylotype IV) and 53 non-RSSC bacterial pathogens on relevant hosts and soil, and 100% diagnostic specificity and sensitivity based on testing inoculated plants and a Euphresco sample panel. The assays presented here represent useful diagnostic tools that can be deployed on-site to aid domestic survey and detection efforts.
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BibTeX
@article{karakkat2026genomeinformed,
title = {{Genome-Informed Loop-Mediated Isothermal Amplification (LAMP) Assays for Detection and Differentiation of Ralstonia pseudosolanacearum Phylotypes I and III}},
author = {Brijesh B. Karakkat and Qi Huang and Schyler O. Nunziata and Jarred Yasuhara-Bell},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = oct,
doi = {10.3390/ijms27208976},
url = {https://www.mdpi.com/1422-0067/27/20/8976},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Genome-Informed Loop-Mediated Isothermal Amplification (LAMP) Assays for Detection and Differentiation of Ralstonia pseudosolanacearum Phylotypes I and III
AU - Brijesh B. Karakkat
AU - Qi Huang
AU - Schyler O. Nunziata
AU - Jarred Yasuhara-Bell
PY - 2026
DA - 2026/10/10
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27208976
UR - https://www.mdpi.com/1422-0067/27/20/8976
AB - The Ralstonia solanacearum species complex (RSSC) causes bacterial wilt in a broad range of economically important hosts. The RSSC has been reclassified into three species, of which R. solanacearum sequevars 1 and 2 in phylotype IIB are designated as Select Agents in the U.S. Many rapid tests for “R. solanacearum” target the RSSC and thus cross-react with R. pseudosolanacearum, which is endemic to the U.S. To avoid false positive identifications and unnecessary triggering of Select Agent measures, four portable LAMP-based screening assay variations were developed and validated for specific detection of R. pseudosolanacearum: real-time LAMP assays for detection of R. pseudosolanacearum (1) and differentiation of R. pseudosolanacearum phylotypes I and III in the AmpliFire® Isothermal Fluorometer (2) or Genie® III (3), and a colorimetric LAMP for detection of R. pseudosolanacearum (4). All assays demonstrated high sensitivity (10–50 colony-forming units (CFU)/reaction) and specificity; 100% analytical specificity and selectivity based on 177 RSSC isolates (81 phylotype I, 70 phylotype II, 15 phylotype III and 11 phylotype IV) and 53 non-RSSC bacterial pathogens on relevant hosts and soil, and 100% diagnostic specificity and sensitivity based on testing inoculated plants and a Euphresco sample panel. The assays presented here represent useful diagnostic tools that can be deployed on-site to aid domestic survey and detection efforts.
SN - 1422-0067
ER -