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Highly Targeted Detection of Priority Phytopathogen : From Obtaining Polyclonal Antibodies to Development and Approbation of Enzyme-Linked Immunoassay and Lateral Flow Immunoassay. | LitMetric

is a bacterial phytopathogen that causes soft and black rot and actively spreads worldwide. Our study is the first development of immunoassays for detecting . We immunized rabbits and obtained serum with an extremely high titer (1:10). Isolated polyclonal antibodies were tested by enzyme-linked immunosorbent assay (ELISA) using 18 closely related strains and 5 non-related bacterial pathogens. No cross-reactivity was found concerning the tested pathogens. The ELISA of was developed in a double-antibody sandwich format with a detection limit of 1.5 × 10 cells/mL. A lateral flow immunoassay (LFIA) for was also developed in a double-antibody sandwich format with a detection limit of 1 × 10 cells/mL. The results of cells testing with LFIA in plant matrix showed a high correlation (R = 0.932) between concentrations of added and revealed cells. When testing potato seed material, ELISA and LFIA confirmed 75 and 66% of positive samples according to real-time PCR, respectively. For negative samples, ELISA showed 84% coincidence, and LFIA coincided with PCR for 89% of samples. Thus, the developed immunoassays can be used to evaluate plant material in poorly equipped conditions or under field testing.

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http://dx.doi.org/10.3390/microorganisms12122436DOI Listing

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