Use of ICELISA to detect Nosema perny

Author of the article:JIANG YiRen1,2WANG BoYang1SUN Ying1WANG Yong2SHI ShengLin1YANG RuiSheng1DUAN YuXi2**QIN Li1**

Author's Workplace：1.College of Bioscience & Biotechnology, Shenyang Agricultural University, Liaoning Engineering &Technology Research Center for Insects Resources, Shenyang 110866, China; 2.College of Plant Protection, Shenyang Agricultural University, Shenyang110866, China

Key Words：Nosema pernyi, ELISA, polyclonal antibody, detection

Abstract：Microsporidiosis is the only quarantinable disease of Antheraea pernyi. Nosema pernyi is the lethal pathogen of microsporidiosis. Therefore, the detection of the spores of N. pernyi is important to prevent and treat this disease. In this paper, the effectiveness of an indirect competitive ELISA for detecting the spores of N. pernyi in A. pernyi was studied by preparing the polyclonal antibody for N. pernyi. The polyclonal antibody against N. pernyi was prepared by immunizing rabbits using a suspension containing spores of N. pernyi. The titre and antibody concentration were 1∶104 and 3 mg·mL-1 respectively, the antibody mainly contained two protein straps with molecular weights of 50 ku and 25 ku, respectively. The resultant polyclonal antibody can be kept for further study. The optimal antigen concentration for the ICELISA was 20 μg·mL-1 spore wall protein of N. pernyi, the optimal polyclonal antibody concentration was 1∶102, HRPIgG optimal concentration was 1∶5×104 and the sensitivity of the method was 1.6×105 spores·mL-1. The method had some value in the detection of N. pernyi.