title: Evaluation of recombinant multi-epitope proteins for diagnosis of goat schistosomiasis by enzyme-linked immunosorbent assay.
abstract: BACKGROUND
  Schistosomiasis is a huge threat to human and animal health. Apart from bovines, goats play an important role in the transmission of schistosomiasis in some endemic areas of China. An accessible, quality-assured goat schistosomiasis diagnostic technique is needed. Recently, our laboratory identified two recombinant diagnostic antigens, SjPGM and SjRAD23 via an immuno-proteomic method. The application of these two recombinant antigens to develop a higher sensitivity and specificity technique for the sheep schistosomiasis diagnosis is urgently needed.
  
  
  METHODS
  Epitopes of SjPGM and SjRAD23 were predicted and three polypeptides, two from SjRAD23 and one from SjPGM, were selected. Recombinant plasmids containing two to three DNA sequences encoding predicted polypeptides or large hydrophilic region of Sj23 (LHD-Sj23) were constructed and expressed. Eight recombinant schistosome antigens including four multi-epitope proteins and four recombinant single-molecule antigens as well as SEA, were assessed by ELISA in 91 sera from schistosome-infected goats, 44 sera from non-infected goats, 37 sera from Orientobilharzia-infected goats, and 12 from Haemonchus contortus-infected goats.
  
  
  RESULTS
  ELISA tests showed that three multi-epitope proteins had higher sensitivity than the four single-molecule antigens (rSjRAD23, rSjPGM, rBSjRAD23-1, rBSj23) and the multi-epitope protein rBSjPGM-BSjRAD23-1-BSj23 had the highest sensitivity (97.8 %, 89/91) and maintained good specificity (100 %, 44/44) as well as low cross-reactivity with haemonchosis (8.33 %, 3/12) and orientobilharziasis (13.51 %, 5/37) in the diagnosis of goat schistosomiasis. In contrast, when SEA was applied as a diagnosis antigen, it had 100 % (91/91) sensitivity, 75 % (33/44) specificity, 25 and 83.78 % cross-reactivity with haemonchosis (3/12) and orientobilharziasis (31/37), respectively.
  
  
  CONCLUSIONS
  The application of recombinant multi-epitope proteins may increase the sensitivity of diagnosis technique and retain high specificity of single-molecule antigens for schistosomiasis, and the recombinant antigen rBSjPGM-BSjRAD23-1-BSj23 has the potential to be used as a diagnosis antigen for goat schistosomiasis.
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. 842684360@qq.com.|College of Life and Environmental Science, Shanghai Normal University, Shanghai, China. 842684360@qq.com.
authors.forename: Chao
authors.initials: C
authors.lastname: Lv
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. hongyang_7@126.com.
authors.forename: Yang
authors.initials: Y
authors.lastname: Hong
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. fuzhiqiang@shvri.ac.cn.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Fu
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. luke@shvri.ac.cn.
authors.forename: Ke
authors.initials: K
authors.lastname: Lu
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. 330112590@qq.com.
authors.forename: Xiaodan
authors.initials: X
authors.lastname: Cao
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. 1205641203@qq.com.
authors.forename: Tao
authors.initials: T
authors.lastname: Wang
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. zcg@shvri.ac.cn.
authors.forename: Chuangang
authors.initials: C
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. mickeyli7102@aliyun.com.
authors.forename: Hao
authors.initials: H
authors.lastname: Li
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. piaooluo@163.com.
authors.forename: Rui
authors.initials: R
authors.lastname: Xu
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. 993361216@qq.com.
authors.forename: Bingguang
authors.initials: B
authors.lastname: Jia
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. 760537541@qq.com.
authors.forename: Qian
authors.initials: Q
authors.lastname: Han
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. 657392996@qq.com.|College of Life and Environmental Science, Shanghai Normal University, Shanghai, China. 657392996@qq.com.
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Dou
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. 857281247@qq.com.|College of Life and Environmental Science, Shanghai Normal University, Shanghai, China. 857281247@qq.com.
authors.forename: Yuanxi
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. 561656239@qq.com.
authors.forename: Zuhang
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. 1393707983@qq.com.
authors.forename: Jinli
authors.initials: J
authors.lastname: Zai
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. 279351182@qq.com.
authors.forename: Jintao
authors.initials: J
authors.lastname: Feng
authors.affiliation: Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China. jjlin@shvri.ac.cn.|Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, China. jjlin@shvri.ac.cn.
authors.forename: Jiaojiao
authors.initials: J
authors.lastname: Lin
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood*; D000947:Antigens, Helminth / Q000235:genetics / Q000276:immunology*; D019411:Clinical Laboratory Techniques / Q000379:methods*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D015511:Goat Diseases / Q000175:diagnosis*; D006041:Goats; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012552:Schistosomiasis / Q000175:diagnosis / Q000662:veterinary*; D012680:Sensitivity and Specificity; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D000939:Epitopes; D011994:Recombinant Proteins