Loop-mediated isothermal amplification (LAMP) assay coupled with gold nanoparticles for colorimetric detection of Trichoderma spp. in Agaricus bisporus cultivation substrates.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
05 Jul 2024
05 Jul 2024
Historique:
received:
14
01
2024
accepted:
25
06
2024
medline:
6
7
2024
pubmed:
6
7
2024
entrez:
5
7
2024
Statut:
epublish
Résumé
One of the significant challenges in organic cultivation of edible mushrooms is the control of invasive Trichoderma species that can hinder the mushroom production and lead to economic losses. Here, we present a novel loop-mediated isothermal amplification (LAMP) assay coupled with gold nanoparticles (AuNPs) for rapid colorimetric detection of Trichoderma spp. The specificity of LAMP primers designed on the tef1 gene was validated in silico and through gel-electrophoresis on Trichoderma harzianum and non-target soil-borne fungal and bacterial strains. LAMP amplification of genomic DNA templates was performed at 65 °C for only 30 min. The results were rapidly visualized in a microplate format within less than 5 min. The assay is based on salt-induced aggregation of AuNPs that is being prevented by the amplicons produced in case of positive LAMP reaction. As the solution color changes from red to violet upon nanoparticle aggregation can be observed with the naked eye, the developed LAMP-AuNPs assay can be easily operated to provide a simple initial screening for the rapid detection of Trichoderma in button mushroom cultivation substrate.
Identifiants
pubmed: 38969670
doi: 10.1038/s41598-024-65971-9
pii: 10.1038/s41598-024-65971-9
doi:
Substances chimiques
Gold
7440-57-5
DNA, Fungal
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
15539Subventions
Organisme : European Commission
ID : IPANEMA project, N° 872662
Organisme : Ministry of Science, Technological Development, and Innovations of the Republic of Serbia
ID : 451-03-66/2024-03/200358
Organisme : Agence Nationale de la Recherche
ID : ANR-21-CE21-009 SIENA
Informations de copyright
© 2024. The Author(s).
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