Biotechnology Bulletin ›› 2018, Vol. 34 ›› Issue (6): 79-83.doi: 10.13560/j.cnki.biotech.bull.1985.2017-1012

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Preparation of Polyclonal Antibody Against Ratoon Stunting Disease and Its Immunomagnetic Beads

GUO Ying1 ,WANG Wen-hua1 ,LIU Li-qing1 ,HU Min2   

  1. 1. Fujian Key Laboratory of Subtropical Plant Physiology and Biochemistry,Fujian Institute of Subtropical Botany,Xiamen 361006;
    2. Institute of Chemical Technology,Huaqiao University,Xiamen 361021
  • Received:2017-11-29 Online:2018-06-26 Published:2018-07-03

Abstract: This study is to isolate sugarcane Lxx by preparing Lxx immunomagnetic beads(IMB)for the further study the interaction between pathogen and sugarcane. The pathogen of Lxx was isolated from the RSD PCR-verified infected sugarcane for the preparation of polyclonal antibody. Then the Lxx polyclonal antibody was coupled with magnetic Fe3O4 nano particle for preparing IMB. The isolates were identified as Lxx according to the morphological characteristics of colony and the PCR results. The fine polyclonal antibody was obtained by immunizing rabbits with Lxx. The titer of the polyclonal antibody against Lxx was greater than 204,800 by indirect ELISA. The appearance of the IMB was globular,and the particulates were 125-268 nm in diameter. The isolates with high affinity for Lxx in the culture media can be used for PCR reaction. The successful preparation of Lxx IMB can rapidly isolate and enrich Lxx,which provides a convenient and efficient technique for the detection of pathogenic bacteria under the interaction of Lxx and sugarcane.

Key words: ratoon stunting disease, pathogenic bacterium Lxx, polyclonal antibody, Lxx-IMB