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Article

Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies

1
Centro de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), CV-315 km 10.7, 46113 Moncada, Valencia, Spain
2
INGENASA, Hermanos García Noblejas 39, 28037 Madrid, Spain
3
PLANT PRINT Diagnòstics SL, Mayor 25, 46610 Guadassuar, Valencia, Spain
*
Author to whom correspondence should be addressed.
Agronomy 2021, 11(1), 48; https://doi.org/10.3390/agronomy11010048
Received: 24 November 2020 / Revised: 21 December 2020 / Accepted: 22 December 2020 / Published: 28 December 2020
(This article belongs to the Special Issue Diagnosis, Population Biology and Management of Vascular Diseases)
Monoclonal antibodies (MAb) specific to Xylella fastidiosa were obtained through hybridoma technology using heat-treated somatic O antigens from LMG 17159strain. Ten stable hybrydoma clones secreting MAb were selected and their isotype was determined. The MAbs 2G1/PPD, IgG1 showed specificity for X. fastidiosa, detecting all the analyzed strains representing different subspecies, STs and hosts. Polyclonal antibodies (PAb) against X. fastidiosa were also produced and antiserum 17159-O/IVIA was selected for the highest titre and its excellent detection capability. MAb 2G1/PPD was tested against strain IVIA 5235 in PBS and in spiked raw extract samples from almond, olive, citrus, and other hosts and its sensitivity by DAS-ELISA was 104 CFU mL−1. The MAb also reacted with high affinity and avidity against X. fastidiosa by DASI-ELISA and Tissue print-ELISA. The diagnostic parameters of DAS-ELISA based on MAb were calculated and compared with the gold standard real-time PCR. The diagnostic specificity of MAb2G1/PPD was 100%, the diagnostic sensitivity was 88.5% compared to Harper’s real-time PCR and 89.9% compared to Francis’ real-time PCR. The agreement between the techniques was almost perfect according to the estimated Cohen’s kappa-index, even in symptomless almond trees. The developed immunological techniques represent sustainable and low-cost analysis tools, based on specific, homogeneous, and well-characterized MAbs, which can be obtained in unlimited quantities in a reproducible way and constitute a guarantee for the standardization of commercial kits. They are a valuable option within a polyphasic strategy for the detection of X. fastidiosa. View Full-Text
Keywords: DAS-ELISA; DASI-ELISA; tissue print-ELISA; diagnostic parameters; Cohen’s kappa index; sustainable diagnosis DAS-ELISA; DASI-ELISA; tissue print-ELISA; diagnostic parameters; Cohen’s kappa index; sustainable diagnosis
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MDPI and ACS Style

Gorris, M.T.; Sanz, A.; Peñalver, J.; López, M.M.; Colomer, M.; Marco-Noales, E. Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies. Agronomy 2021, 11, 48. https://doi.org/10.3390/agronomy11010048

AMA Style

Gorris MT, Sanz A, Peñalver J, López MM, Colomer M, Marco-Noales E. Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies. Agronomy. 2021; 11(1):48. https://doi.org/10.3390/agronomy11010048

Chicago/Turabian Style

Gorris, María T., Antonio Sanz, Javier Peñalver, María M. López, Mario Colomer, and Ester Marco-Noales. 2021. "Detection and Diagnosis of Xylella fastidiosa by Specific Monoclonal Antibodies" Agronomy 11, no. 1: 48. https://doi.org/10.3390/agronomy11010048

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