Naked-Eye Molecular Testing for the Detection of <i>Xylella fastidiosa</i> in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP

Naked-Eye Molecular Testing for the Detection of <i>Xylella fastidiosa</i> in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP

<i>Xylella fastidiosa</i> (<i>Xf</i>) is a quarantine pathogen heavily affecting economically important crops worldwide. Different sequence types (STs) belonging to <i>Xf</i> subspecies are present in various areas of Spain, including the Balearic Islands, and cau...

Full description

Saved in:
Bibliographic Details
Main Authors: Amoia Serafina Serena, Ana Falcón-Piñeiro, Milica Pastar, José Manuel Garcìa-Madero, Nicoletta Contaldo, Mikael Muegge, Stéphane Compant, Pasquale Saldarelli, Angelantonio Minafra
Format: Article
Language:English
Published: MDPI AG 2025-01-01
Series:Applied Sciences
Subjects:
almond leaf scorch
<i>Xylella fastidiosa</i>
diagnosis
colorimetric LAMP
point of care
Online Access:https://www.mdpi.com/2076-3417/15/2/739
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:<i>Xylella fastidiosa</i> (<i>Xf</i>) is a quarantine pathogen heavily affecting economically important crops worldwide. Different sequence types (STs) belonging to <i>Xf</i> subspecies are present in various areas of Spain, including the Balearic Islands, and cause the almond leaf scorch disease (ALSD) in <i>Prunus</i> spp. The increased demand for rapid tests for early detection of the pathogen should enforce strict containment measures. Molecular detection through isothermal amplification reactions enables simplified instrumentation and the use of raw nucleic acid extracts. Colorimetric loop-mediated isothermal amplification (cLAMP) was applied to rapidly detect <i>Xf</i> in naturally infected almonds on Mallorca Island (Spain), using a quick crude sap extraction without DNA purification. Following tissue homogenization, an alkaline treatment for target DNA extraction was conducted before the cLAMP test. The cLAMP assay was able to detect up to 100 CFU/mL of the <i>Xf</i> bacterial suspension diluted in healthy almond sap. The same crude extracts used in the cLAMP test were also tested by qPCR. An overall positive agreement of about 47% was observed between the results of the two techniques, while a decrease in cLAMP sensitivity was evident as the bacterial titer declined in infected plants over Cq > 26–27. This study shows the potential of the cLAMP application as a rapid and low-cost point-of-care diagnostic method for the timely monitoring of <i>Xf</i> directly in the field.
ISSN:2076-3417

Similar Items