Evidence map›Paper›PMID 36357509›Full record

ArticleScientific reports2022

Loop-mediated isothermal amplification (LAMP) assay for specific and rapid detection of Dickeya fangzhongdai targeting a unique genomic region.

Anuhea DeLude, Riley Wells, Sherine Boomla, Shu-Cheng Chuang, Frank Urena, Aaron Shipman, Noelle Rubas, Donna Lee Kuehu, Buster Bickerton, Taylor Peterson and 12 more

Abstract read
In one paragraph

Article in Scientific reports, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed.

  1. Frontiers in microbiology · 2024
    Article
  2. Retrospective survey ofFrontiers in microbiology · 2023
    Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

22 authors.

Anuhea DeLude *Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI, USA.
Riley Wells *Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, USA.
Sherine Boomla *Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, USA.
Shu-Cheng Chuang *Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI, USA.
Frank Urena *Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, USA.
Aaron Shipman *Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI, USA.
Noelle Rubas *Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, USA.
Donna Lee Kuehu *Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, USA.
Buster Bickerton *Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, USA.
Taylor Peterson *Department of Human Nutrition, Food and Animal Sciences, University of Hawaii at Manoa, Honolulu, HI, USA.
Shefali DobhalDepartment of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI, USA.
Dario ArizalaDepartment of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI, USA.
Diksha KlairDepartment of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI, USA.
Francisco Ochoa-CoronaInstitute for Biosecurity & Microbial Forensics, Oklahoma State University, Stillwater, OK, USA.
Md Emran AliDepartment of Plant Pathology, University of Georgia, Tifton, GA, USA.
Jenee OdaniDepartment of Human Nutrition, Food and Animal Sciences, University of Hawaii at Manoa, Honolulu, HI, USA.
Jon-Paul BinghamDepartment of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, USA.
Daniel M JenkinsDepartment of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, USA.
Jacqueline FletcherInstitute for Biosecurity & Microbial Forensics, Oklahoma State University, Stillwater, OK, USA.
James P StackDepartment of Plant Pathology, Kansas State University, Manhattan, KS, USA.
Anne M AlvarezDepartment of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI, USA.
Mohammad ArifDepartment of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI, USA. arif@hawaii.edu.

Funding

UH Hilo COP A&RP20GM103466 · NIGMS · UNIVERSITY OF HAWAII AT MANOA · PI HELEN C TURNER · 2012 to 2026
$60.3M
NIGMS NIH HHS P20 GM103466
6 · The paper itself

Abstract

Dickeya fangzhongdai, a bacterial pathogen of taro (Colocasia esculenta), onion (Allium sp.), and several species in the orchid family (Orchidaceae) causes soft rot and bleeding canker diseases. No field-deployable diagnostic tool is available for specific detection of this pathogen in different plant tissues. Therefore, we developed a field-deployable loop-mediated isothermal amplification (LAMP) assay using a unique genomic region, present exclusively in D. fangzhongdai. Multiple genomes of D. fangzhongdai, and other species of Dickeya, Pectobacterium and unrelated genera were used for comparative genomic analyses to identify an exclusive and conserved target sequence from the major facilitator superfamily (MFS) transporter gene region. This gene region had broad detection capability for D. fangzhongdai and thus was used to design primers for endpoint PCR and LAMP assays. In-silico validation showed high specificity with D. fangzhongdai genome sequences available in the NCBI GenBank genome database as well as the in-house sequenced genome. The specificity of the LAMP assay was determined with 96 strains that included all Dickeya species and Pectobacterium species as well as other closely related genera and 5 hosts; no false positives or false negatives were detected. The detection limit of the assay was determined by performing four sensitivity assays with tenfold serially diluted purified genomic DNA of D. fangzhongdai with and without the presence of crude host extract (taro, orchid, and onion). The detection limit for all sensitivity assays was 100 fg (18-20 genome copies) with no negative interference by host crude extracts. The assays were performed by five independent operators (blind test) and on three instruments (Rotor-Gene, thermocycler and dry bath); the assay results were concordant. The assay consistently detected the target pathogen from artificially inoculated and naturally infected host samples. The developed assay is highly specific for D. fangzhongdai and has applications in routine diagnostics, phytosanitary and seed certification programs, and epidemiological studies.

Indexed as

OrchidaceaePectobacteriumDickeyaEnterobacteriaceaeGenomicsMolecular Diagnostic TechniquesNucleic Acid Amplification TechniquesSensitivity and Specificity

Identifiers

PMID36357509
PMCPMC9649655

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.