Dissertation > Excellent graduate degree dissertation topics show

Preparation of the Monoclonal Antibody Against S-LPS and Development of Competitive ELISA for Detection of Antibodies Against Brucella

Author: WangJiaLan
Tutor: BuZhiGao
School: Chinese Academy of Agricultural Sciences
Course: Preventive Veterinary Medicine
Keywords: Brucella smooth lipopolysaccharide (S-LPS) competitive ELISA (C-ELISA) monoclonal antibody(McAb)
CLC: S854.43
Type: Master's thesis
Year: 2009
Downloads: 207
Quote: 0
Read: Download Dissertation

Abstract


The serological responses following infection with smooth Brucella species are directed predominantly against the smooth lipopolysaccharide (S-LPS). Thus, in humans as well as in animals, the diagnosis of brucellosis is usually based on the detection of specific antibodies against S-LPS.The O polysaccharides of the smooth lipopolysaccharide is a homopolymer of 4,6-dideoxy-4-formamidoct-D-mannopyranose residues. Up to now,There were four types of antigen has been recognized : The A antigen, the M antigen, the common antigen (C) specificity for smooth Brucella and the common antigen crossreacting with Y.enterocolitica O:9 (C/Y).A problematic aspect of traditional serological detection was false-positive serological reactions (FPSRs). These FPSRs find their origins in the cross-reactivity observed between the S-LPS of Brucella species and the LPSs of some other Gram-negative bacterias, In particular Y.enterocolitica O:9. Whether the traditional detections such as tube agglutination test, rose bengal plate test and complement fixation test, or indirect ELISA with S-LPS as coating antigen, the FPSRs can not be ruled out .Based on the problem of cross-reaction may be solved by the monoclonal antibody aginst C epitopes of Brucella can solution, The B.melitensis M5-90 were inactivated to immune BALB/c mice, hybridomas were obtained by standard cell fusion techniques in the study. Extraction, Purification and Identification of LPS from Brucella vaccine strains M5-90, S-19 and Y.enterocolitica O:9 and Two cell strains secreting monoclonal antibody (McAb) against S-LPS, named 4G6 and 16C5, were screened by I-ELISA using three kinds of LPS .Specificity analysis showed that McAb 4G6 has no cross reaction with Gram-negative bacterias except Y.enterocolitica O:9 while McAb 16C5 with high sensitivity has no cross reaction with all of other bacterias Involved . Ascites of McAb 16C5 was purified by protein G and used in competitive ELISA (C-ELISA) as detect antibodies to develop the method for detection of antibodies against Brucella. A large number of clinical samples was detected to optimize reaction conditions and evaluat the detection methods by Compared with the rose bengal plate agglutination test, IDEXX indirect ELISA Kit, Sweden Svanova competition ELISA Kit and indirect ELISA kit produce by OIE recommended technology .Serum samples from cattle (n=507) were tested by the rose Bengal plate test (RBT) and IDEXX indirect enzyme-linked immunosorbent assay (IELISA) kits and the data demonstrated that the coincidence between C-ELISA and RBT ,C-ELISA and IDEXX IELISA kit was 84.7% and 91.2% respectively. One hundred and fourty-two of the 507 serum were tested and the results showed that the coincidence between C-ELISA and PINGHE IELISA , C-ELISA and Svanova C-ELISA was 94.4% and 97.2% respectively. The results of the C-ELISA are in good agreement (κ>0.5) with those of RBT, IDEXX IELISA kit, PINGHE IELISA kit, Svanova C-ELISA kit, which concluded that the C-ELISA has good reliability. Thus, the CELISA was a valuable asset to the diagnosis of brucellosis and could eliminate false-positive serological reactions that caused by Y.enterocolitica O:9, E. coli O:157 and so on.

Related Dissertations

  1. Identification of B.melitensis 16M Protein Immunodominant Antigens,S852.61
  2. Studies on Dynamic Regularity of Antibody Against G. Anatis and Development of Hybridoma Cell Lines Secreting Monoclonal Antibodies Against G. Anatis Lps,S858.32
  3. Epidemiological Investigation of Brucellosis in Livestock and Molecule Typing of Brucella Based on Single Nucleotide Polymorphism (SNP) Analysis,S855.12
  4. The Effectiveness of GSL in Brucella Ovis 019 Strain Infecting Mouse Macrophage,S852.61
  5. Preparation of Bovine Brucellosis Diagnostic Antigen by Using a Phage Display Library and Its Application in Diagnosis,S858.23
  6. Development and Application of Monoclonal Antibodies Against Zearalenone,S852.43
  7. Studies on the Preparation for Anti-Pb～(2+) Monoclonal Antibody (McAb) and the Method of DTC Chelating ELISA Detection for Hg～(2+),X171
  8. Preparation and Preliminary Characterization of Bispecific Antibody Against Anti-human Laryngeal Carcinoma and Anti-VEGF,R739.65
  9. Study on Rapid Immunology Screening Methods of Fumonisins in Corn,R155.5
  10. Study on the Function of FTH1 in the Course of Brucella Ovis 019 Infecting RAW264.7 Macrophages,S852.61
  11. Study on the Molecular Mechanism of Pathogenesis for Brucella Ovis,S858.26
  12. Preparation of Monoclonal Antibody to a Novel Hepatocellular Carcinoma Marker MXR7 and Clinical Application,R392
  13. Expression, Identification of Bioactive Characterization and Antibacterial Kinetics of Brucellaphage Tbilisi Lysin Gene,R378
  14. Construction of bp26 Gene Deletion Mutant in B.Melitensis Vaccine Strain M5-90 and Evaluation of Safety and Immunogenicity,S852.5
  15. Differential Diagnosis of Brucellosis Marked Vaccine (M5-90-26) and Immunogenicity Analysis of bp26 Protein,S855.12
  16. Preliminary Application and Establishment of Monoclonal Antibody Based Latex Agglutination Test for Detection of B.abortus and B.melitensis,S854.43
  17. Brucella outer membrane protein Omp39 and Omp14 prokaryotic expression and Immunogenicity,S852.5
  18. Isolation, Identification and Characterization of a Phenol-degrading Strain,X172
  19. Detection and Molecular Typing of Milk Brucella Abortus in Cows,S854.43
  20. Research on the GICA Dipsticks of Brucella Strain Specificity Antigens and Antibodies and Application,S854.43

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Veterinary clinical medicine > Veterinary diagnostics > Laboratory diagnostic method
© 2012 www.DissertationTopic.Net  Mobile