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...
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2025-01-01
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| author | Amoia Serafina Serena Ana Falcón-Piñeiro Milica Pastar José Manuel Garcìa-Madero Nicoletta Contaldo Mikael Muegge Stéphane Compant Pasquale Saldarelli Angelantonio Minafra |
| author_facet | Amoia Serafina Serena Ana Falcón-Piñeiro Milica Pastar José Manuel Garcìa-Madero Nicoletta Contaldo Mikael Muegge Stéphane Compant Pasquale Saldarelli Angelantonio Minafra |
| author_sort | Amoia Serafina Serena |
| collection | DOAJ |
| description | <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. |
| format | Article |
| id | doaj-art-4063c3fb68ec448487deeb48f0e7d08c |
| institution | Kabale University |
| issn | 2076-3417 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
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| series | Applied Sciences |
| spelling | doaj-art-4063c3fb68ec448487deeb48f0e7d08c2025-01-24T13:20:39ZengMDPI AGApplied Sciences2076-34172025-01-0115273910.3390/app15020739Naked-Eye Molecular Testing for the Detection of <i>Xylella fastidiosa</i> in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMPAmoia Serafina Serena0Ana Falcón-Piñeiro1Milica Pastar2José Manuel Garcìa-Madero3Nicoletta Contaldo4Mikael Muegge5Stéphane Compant6Pasquale Saldarelli7Angelantonio Minafra8Institute for Sustainable Plant Protection (IPSP)—National Research Council, 70126 Bari, ItalyDMC Research Center, 18620 Granada, SpainAIT Austrian Institute of Technology, Center for Health and Bioresources, Bioresources Unit, 3430 Tulln, AustriaDMC Research Center, 18620 Granada, SpainInstitute for Sustainable Plant Protection (IPSP)—National Research Council, 70126 Bari, ItalyRTDS Group, Lerchengasse 25/2-3, 1080 Vienna, AustriaAIT Austrian Institute of Technology, Center for Health and Bioresources, Bioresources Unit, 3430 Tulln, AustriaInstitute for Sustainable Plant Protection (IPSP)—National Research Council, 70126 Bari, ItalyInstitute for Sustainable Plant Protection (IPSP)—National Research Council, 70126 Bari, Italy<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.https://www.mdpi.com/2076-3417/15/2/739almond leaf scorch<i>Xylella fastidiosa</i>diagnosiscolorimetric LAMPpoint of care |
| spellingShingle | Amoia Serafina Serena Ana Falcón-Piñeiro Milica Pastar José Manuel Garcìa-Madero Nicoletta Contaldo Mikael Muegge Stéphane Compant Pasquale Saldarelli Angelantonio Minafra Naked-Eye Molecular Testing for the Detection of <i>Xylella fastidiosa</i> in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP Applied Sciences almond leaf scorch <i>Xylella fastidiosa</i> diagnosis colorimetric LAMP point of care |
| title | Naked-Eye Molecular Testing for the Detection of <i>Xylella fastidiosa</i> in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP |
| title_full | Naked-Eye Molecular Testing for the Detection of <i>Xylella fastidiosa</i> in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP |
| title_fullStr | Naked-Eye Molecular Testing for the Detection of <i>Xylella fastidiosa</i> in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP |
| title_full_unstemmed | Naked-Eye Molecular Testing for the Detection of <i>Xylella fastidiosa</i> in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP |
| title_short | Naked-Eye Molecular Testing for the Detection of <i>Xylella fastidiosa</i> in Mallorca (Balearic Island) Almond Orchards by Colorimetric LAMP |
| title_sort | naked eye molecular testing for the detection of i xylella fastidiosa i in mallorca balearic island almond orchards by colorimetric lamp |
| topic | almond leaf scorch <i>Xylella fastidiosa</i> diagnosis colorimetric LAMP point of care |
| url | https://www.mdpi.com/2076-3417/15/2/739 |
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