title: Production and application of polyclonal and monoclonal antibodies against Spiroplasma eriocheiris.
abstract: A new species of spiroplasma, Spiroplasma eriocheiris (S. eriocheiris), was identified as a lethal pathogen of tremor disease (TD) in Chinese mitten crab recently. In order to acquire appropriate biological and diagnostic tools for characterizing this newly discovered pathogen, 5 monoclonal antibodies (mAbs) and a polyclonal antibody (pAb) against S. eriocheiris were produced. Among the mAbs, 6F5, 7C8 and 12H5 lead to the deformation of S. eriocheiris. A peptide sequence, YMRDMQSGLPRY was identified as a mimic motif of MreB that is the cell shape determining protein of S. eriocheiris interacting with 3 mAbs. Furthermore, a double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for detection of S. eriocheiris was established using the mAb and pAb we prepared. It detected as low as 0.1 μg/mL of S. eriocheiris. No cross-reaction was observed with three other common bacteria (Pseudomonas aeruginosa, Escherichia coli, and Bacillus subtilis) and the hemolymph samples of healthy Eriocheir sinensis. Collectively, our results indicated that the mAbs and pAb we prepared could be used in the analysis of S. eriocheiris membrane proteins mimotope and development of a diagnostic kit for S. eriocheiris infections.
authors.affiliation: Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, Department of Microbiology and Immunology, Medical School, Southeast University, 87 Dingjiaqiao Road, Nanjing 210009, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, Department of Microbiology and Immunology, Medical School, Southeast University, 87 Dingjiaqiao Road, Nanjing 210009, China.
authors.forename: Haixun
authors.initials: H
authors.lastname: Bao
authors.affiliation: Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, Department of Microbiology and Immunology, Medical School, Southeast University, 87 Dingjiaqiao Road, Nanjing 210009, China.
authors.forename: Fengqin
authors.initials: F
authors.lastname: Miao
authors.affiliation: Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, Department of Microbiology and Immunology, Medical School, Southeast University, 87 Dingjiaqiao Road, Nanjing 210009, China.
authors.forename: Yaqin
authors.initials: Y
authors.lastname: Peng
authors.affiliation: Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, Department of Microbiology and Immunology, Medical School, Southeast University, 87 Dingjiaqiao Road, Nanjing 210009, China.
authors.forename: Yuqing
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Jiangsu Key Laboratory for Biodiversity &Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210023, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Gu
authors.affiliation: Jiangsu Key Laboratory for Biodiversity &Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210023, China.
authors.forename: Qingguo
authors.initials: Q
authors.lastname: Meng
authors.affiliation: Jiangsu Key Laboratory for Biodiversity &Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210023, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Wang
authors.affiliation: Key Laboratory of Developmental Genes and Human Disease, Ministry of Education, Department of Microbiology and Immunology, Medical School, Southeast University, 87 Dingjiaqiao Road, Nanjing 210009, China.
authors.forename: Jianqiong
authors.initials: J
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay; D019151:Peptide Library; D011817:Rabbits; D011995:Recombination, Genetic / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013149:Spiroplasma / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D019151:Peptide Library