title: [Identification of serological antigens in excretory-secretory antigens of Trichinella spiralis muscle larvae].
abstract: OBJECTIVE
  To isolate and identify serological antigens in the excretory-secretory antigens of Trichinella spiralis muscle larvae by the combination of co-immunoprecipitation and mass spectrometric technology.
  
  
  METHODS
  The serum IgG of New Zealand rabbits infected with Trichinella spiralis was isolated by ammonium sulfate precipitation. Muscle larvaes were isolated from the infected muscle, and then purified and cultured to collect excretory-secretory antigens. Serological antigens in excretory-secretory antigens were isolated by co-immunoprecipitation and SDS-PAGE, and analyzed by Western blotting. Moreover, the protein bands in New Zealand rabbit sera infected with Trichinella spiralis were identified by mass spectrometric technology.
  
  
  RESULTS
  Indirect ELISA showed that the titer of serum antibody of New Zealand rabbits infected with Trichinella spiralis was 1:6400. The rabbit serum IgG was effectively isolated by ammonium sulfate precipitation. A total of four clear protein bands of the excretory-secretory antigens of Trichinella spiralis were obtained by electrophoresis. Among them, three clear protein bands with relative molecular mass (Mr) being 40 kDa, 50 kDa and 83 kDa were recognized by the rabbit sera infected with Trichinella spiralis but not recognized by the normal rabbit sera. The obtained four protein molecules were confirmed as serine protease, specific serine protease of muscle larvae, 43 kDa secreted glycoprotein and 53 kDa excretory-secretory antigen.
  
  
  CONCLUSIONS
  Four proteins were obtained from the excretory-secretory antigens of Trichinella spiralis muscle larvae by combination of co-immunoprecipitation and mass spectrometric technique analysis, which provided new sources and insights for the diagnosis and vaccine candidates of Trichinellosis.
authors.affiliation: Departement of Parasitology, Zunyi Medical College, Zunyi, Guizhou 563003, China.
authors.forename: Xuegui
authors.initials: X
authors.lastname: Huang
authors.affiliation: Departement of Parasitology, Zunyi Medical College, Zunyi, Guizhou 563003, China. *Corresponding author, E-mail: helifang2004@sina.com.
authors.forename: Lifang
authors.initials: L
authors.lastname: He
authors.affiliation: Laboratory of Molecular Biology, Medical College Hunan Normal University, Changsha 410013, China.
authors.forename: Shishan
authors.initials: S
authors.lastname: Yuan
authors.affiliation: Departement of Parasitology, Zunyi Medical College, Zunyi, Guizhou 563003, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Liu
authors.affiliation: Departement of Parasitology, Zunyi Medical College, Zunyi, Guizhou 563003, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay; D007074:Immunoglobulin G / Q000097:blood; D047468:Immunoprecipitation; D007814:Larva / Q000276:immunology; D013058:Mass Spectrometry; D008970:Molecular Weight; D011817:Rabbits; D017160:Trichinella spiralis / Q000276:immunology*
chemical_list: D000947:Antigens, Helminth; D007074:Immunoglobulin G