Characterization of Polyclonal Antibody Against Pseudomonas syringae pv. actinidiae Effector Hopz5 by Enzyme Linked Immunoabsorbent Assay

doi:10.13560/j.cnki.biotech.bull.1985.2017-0668

Abstract

Abstract: In order to establish a serological detection method for Pseudomonas syringae pv. actinidiae,we used highly pathogenic PSA3 group specific effector Hopz5 to prepare the polyclonal antibody（PAb-Hopz5）. The characteristics of PAb-Hopz5 was comprehensively analyzed by detecting sample type,specificity,sensitivity and field samples via enzyme linked immunoabsorbent assay . The results showed that PAb-Hopz5 could be used for detecting not only the kiwifruit tissues,but also the PSA of artificial culture;and not only the protein of disease tissue but also the soak solution of tissues could be used for the detection. Besides,the samples of PSA different infection periods could be detected by PAb-Hopz5. No cross-reaction were observed with other testing bacteria,including P. syringae pv. tomato,P. fluorescens and P. putida,etc,indicating that the specificity of PAb-Hopz5 was solid. The minimum detectable concentration of bacterial suspension by ELISA was 3×10⁵CFU/mL,which was significantly lower than that by PCR,but for the detection of disease samples in the field it was better than that by PCR. Overall,the results showed that PAb-Hopz5 could be used as the test reagent for the PSA3 group detection of P. syringae pv. actinidiae,thus it provided a new detection tool for P. syringae pv. actinidiae.

Key words: Pseudomonas syringae pv. Actinidiae, PSA3 group, PAb-Hopz5, serological detection

HU Yue, CHEN Hang, YANG Xun-zhe, LI Qing, YANG Hui. Characterization of Polyclonal Antibody Against Pseudomonas syringae pv. actinidiae Effector Hopz5 by Enzyme Linked Immunoabsorbent Assay[J]. Biotechnology Bulletin, 2018, 34(1): 84-89.

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