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Open AccessBrief Report

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

by

Amoia Serafina Serena

^1,2

,

Ana Falcón-Piñeiro

³

,

Milica Pastar

⁴,

José Manuel Garcìa-Madero

³

,

Nicoletta Contaldo

¹,

Mikael Muegge

⁵,

Stéphane Compant

⁴

,

Pasquale Saldarelli

^1,* and

Angelantonio Minafra

^1,*

¹

Institute for Sustainable Plant Protection (IPSP)—National Research Council, 70126 Bari, Italy

²

Department of Soil, Plant and Food Sciences, University of Bari Aldo Moro, 70126 Bari, Italy

³

DMC Research Center, 18620 Granada, Spain

⁴

AIT Austrian Institute of Technology, Center for Health and Bioresources, Bioresources Unit, 3430 Tulln, Austria

⁵

RTDS Group, Lerchengasse 25/2-3, 1080 Vienna, Austria

^*

Authors to whom correspondence should be addressed.

Appl. Sci. 2025, 15(2), 739; https://doi.org/10.3390/app15020739

Submission received: 23 December 2024 / Revised: 8 January 2025 / Accepted: 10 January 2025 / Published: 13 January 2025

(This article belongs to the Special Issue Emerging Analytical Techniques in Food Industry and Agricultural Products)

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Abstract

Xylella fastidiosa (Xf) is a quarantine pathogen heavily affecting economically important crops worldwide. Different sequence types (STs) belonging to Xf subspecies are present in various areas of Spain, including the Balearic Islands, and cause the almond leaf scorch disease (ALSD) in Prunus 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 Xf 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 Xf 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 Xf directly in the field.

Keywords: almond leaf scorch; Xylella fastidiosa; diagnosis; colorimetric LAMP; point of care

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MDPI and ACS Style

Serena, A.S.; Falcón-Piñeiro, A.; Pastar, M.; Garcìa-Madero, J.M.; Contaldo, N.; Muegge, M.; Compant, S.; Saldarelli, P.; Minafra, A. Naked-Eye Molecular Testing for the Detection of Xylella fastidiosa in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP. Appl. Sci. 2025, 15, 739. https://doi.org/10.3390/app15020739

AMA Style

Serena AS, Falcón-Piñeiro A, Pastar M, Garcìa-Madero JM, Contaldo N, Muegge M, Compant S, Saldarelli P, Minafra A. Naked-Eye Molecular Testing for the Detection of Xylella fastidiosa in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP. Applied Sciences. 2025; 15(2):739. https://doi.org/10.3390/app15020739

Chicago/Turabian Style

Serena, Amoia Serafina, Ana Falcón-Piñeiro, Milica Pastar, José Manuel Garcìa-Madero, Nicoletta Contaldo, Mikael Muegge, Stéphane Compant, Pasquale Saldarelli, and Angelantonio Minafra. 2025. "Naked-Eye Molecular Testing for the Detection of Xylella fastidiosa in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP" Applied Sciences 15, no. 2: 739. https://doi.org/10.3390/app15020739

APA Style

Serena, A. S., Falcón-Piñeiro, A., Pastar, M., Garcìa-Madero, J. M., Contaldo, N., Muegge, M., Compant, S., Saldarelli, P., & Minafra, A. (2025). Naked-Eye Molecular Testing for the Detection of Xylella fastidiosa in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP. Applied Sciences, 15(2), 739. https://doi.org/10.3390/app15020739

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