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Article

Development of a Real-Time Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of Olea Europaea Geminivirus

by
Sofia Bertacca
1,†,
Andrea Giovanni Caruso
1,2,†,
Daniela Trippa
1,
Annalisa Marchese
1,
Antonio Giovino
2,
Slavica Matic
3,
Emanuela Noris
3,
Maria Isabel Font San Ambrosio
4,
Ana Alfaro
4,
Stefano Panno
1,* and
Salvatore Davino
1,*
1
Department of Agricultural, Food and Forest Sciences (SAAF), University of Palermo, Viale delle Scienze, 90128 Palermo, Italy
2
Research Centre for Plant Protection and Certification (CREA), Strada Statale, 113, 90011 Bagheria, Italy
3
Institute for Sustainable Plant Protection, National Research Council (IPSP-CNR), Strada delle Cacce, 73, 10135 Turin, Italy
4
Instituto Agroforestal Mediteráneo, Universitat Politécnica de València (IAM-UPV), Camino de Vera, s/n, 46022 Valencia, Spain
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Plants 2022, 11(5), 660; https://doi.org/10.3390/plants11050660
Submission received: 7 February 2022 / Revised: 25 February 2022 / Accepted: 26 February 2022 / Published: 28 February 2022
(This article belongs to the Special Issue Epidemiology and Control of Plant Diseases)
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Abstract

A real-time loop-mediated isothermal amplification (LAMP) assay was developed for simple, rapid and efficient detection of the Olea europaea geminivirus (OEGV), a virus recently reported in different olive cultivation areas worldwide. A preliminary screening by end-point PCR for OEGV detection was conducted to ascertain the presence of OEGV in Sicily. A set of six real-time LAMP primers, targeting a 209-nucleotide sequence elapsing the region encoding the coat protein (AV1) gene of OEGV, was designed for specific OEGV detection. The specificity, sensitivity, and accuracy of the diagnostic assay were determined. The LAMP assay showed no cross-reactivity with other geminiviruses and was allowed to detect OEGV with a 10-fold higher sensitivity than conventional end-point PCR. To enhance the potential of the LAMP assay for field diagnosis, a simplified sample preparation procedure was set up and used to monitor OEGV spread in different olive cultivars in Sicily. As a result of this survey, we observed that 30 out of 70 cultivars analyzed were positive to OEGV, demonstrating a relatively high OEGV incidence. The real-time LAMP assay developed in this study is suitable for phytopathological laboratories with limited facilities and resources, as well as for direct OEGV detection in the field, representing a reliable method for rapid screening of olive plant material.
Keywords: LAMP; OEGV; Geminiviridae; olive viruses LAMP; OEGV; Geminiviridae; olive viruses

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MDPI and ACS Style

Bertacca, S.; Caruso, A.G.; Trippa, D.; Marchese, A.; Giovino, A.; Matic, S.; Noris, E.; Ambrosio, M.I.F.S.; Alfaro, A.; Panno, S.; et al. Development of a Real-Time Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of Olea Europaea Geminivirus. Plants 2022, 11, 660. https://doi.org/10.3390/plants11050660

AMA Style

Bertacca S, Caruso AG, Trippa D, Marchese A, Giovino A, Matic S, Noris E, Ambrosio MIFS, Alfaro A, Panno S, et al. Development of a Real-Time Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of Olea Europaea Geminivirus. Plants. 2022; 11(5):660. https://doi.org/10.3390/plants11050660

Chicago/Turabian Style

Bertacca, Sofia, Andrea Giovanni Caruso, Daniela Trippa, Annalisa Marchese, Antonio Giovino, Slavica Matic, Emanuela Noris, Maria Isabel Font San Ambrosio, Ana Alfaro, Stefano Panno, and et al. 2022. "Development of a Real-Time Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of Olea Europaea Geminivirus" Plants 11, no. 5: 660. https://doi.org/10.3390/plants11050660

APA Style

Bertacca, S., Caruso, A. G., Trippa, D., Marchese, A., Giovino, A., Matic, S., Noris, E., Ambrosio, M. I. F. S., Alfaro, A., Panno, S., & Davino, S. (2022). Development of a Real-Time Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of Olea Europaea Geminivirus. Plants, 11(5), 660. https://doi.org/10.3390/plants11050660

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