Evidence map›Paper›PMID 39252004›Full record

ArticlePlant methods2024

Rapid real-time quantitative colorimetric LAMP methodology for field detection of Verticillium dahliae in crude olive-plant samples.

Maria Megariti, Alexandra Panagou, Georgios Patsis, George Papadakis, Alexandros K Pantazis, Epaminondas J Paplomatas, Aliki K Tzima, Emmanouil A Markakis, Electra Gizeli

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Article in Plant methods, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 3 papers.

0numbers the graph read from it
0cells of the map it votes in
3citing 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

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3 · Its place in the literature

Who cites it

3 citing papers in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

9 authors.

Maria MegaritiInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, 100 N. Plastira Str., 70013, Heraklion, Greece.
Alexandra PanagouLaboratory of Plant Pathology, Agricultural University of Athens, 75 Iera Odos, 118 55, Athens, Greece.
Georgios PatsisLaboratory of Plant Pathology, Agricultural University of Athens, 75 Iera Odos, 118 55, Athens, Greece.
George PapadakisInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, 100 N. Plastira Str., 70013, Heraklion, Greece.
Alexandros K PantazisInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, 100 N. Plastira Str., 70013, Heraklion, Greece.
Epaminondas J PaplomatasLaboratory of Plant Pathology, Agricultural University of Athens, 75 Iera Odos, 118 55, Athens, Greece.
Aliki K TzimaLaboratory of Plant Pathology, Agricultural University of Athens, 75 Iera Odos, 118 55, Athens, Greece. aliki@aua.gr.
Emmanouil A MarkakisLaboratory of Mycology, Institute of Olive Tree, Subtropical Crops and Viticulture, Hellenic Agricultural Organization DIMITRA, 32A Kastorias Street, Mesa Katsabas, 71307, Heraklion, Crete, Greece. markmanos@hmu.gr.
Electra GizeliInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, 100 N. Plastira Str., 70013, Heraklion, Greece. gizeli@imbb.forth.gr.

Funding

General Secretariat of Research and Technology 2018ΣΕ01300000
6 · The paper itself

Abstract

backgroundVerticilium dahliae is the most important wilt pathogen of olive trees with a broad host range causing devastating diseases currently without any effective chemical control. Traditional detection methodologies are based on symptoms-observation or lab-detection using time consuming culturing or molecular techniques. Therefore, there is an increasing need for portable tools that can detect rapidly V. dahliae in the field.

resultsIn this work, we report the development of a novel method for the rapid, reliable and on-site detection of V. dahliae using a newly designed isothermal LAMP assay and crude extracts of olive wood. For the detection of the fungus, LAMP primers were designed targeting the internal transcribed spacer (ITS) region of the rRNA gene. The above assay was combined with a purpose-built prototype portable device which allowed real time quantitative colorimetric detection of V. dahliae in 35 min. The limit of detection of our assay was found to be 0.8 fg/μl reaction and the specificity 100% as indicated by zero cross-reactivity to common pathogens found in olive trees. Moreover, detection of V. dahliae in purified DNA gave a sensitivity of 100% (Ct < 30) and 80% (Ct > 30) while the detection of the fungus in unpurified crude wood extracts showed a sensitivity of 80% when multisampling was implemented. The superiority of the LAMP methodology regarding robustness and sensitivity was demonstrated when only LAMP was able to detect V. dahliae in crude samples from naturally infected trees with very low infection levels, while nested PCR and SYBR qPCR failed to detect the pathogen in an unpurified form.

conclusionsThis study describes the development of a new real time LAMP assay, targeting the ITS region of the rRNA gene of V. dahliae in olive trees combined with a 3D-printed portable device for field testing using a tablet. The assay is characterized by high sensitivity and specificity as well as ability to operate using directly crude samples such as woody tissue or petioles. The reported methodology is setting the basis for the development of an on-site detection methodology for V. dahliae in olive trees, but also for other plant pathogens.

Indexed as

Defoliating pathotypeField-based detectionLoop mediated isothermal amplificationMolecular diagnosticsSoilborne pathogensVerticillium wilt

Identifiers

PMID39252004
PMCPMC11386372

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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.