Dissertation > Excellent graduate degree dissertation topics show

Preparation and Preliminary Application of Immune Colloidal Gold Strip for Diagnosis of Goose Paramyxovirus Disease

Author: LvZiHua
Tutor: DingZhuang
School: Jilin University
Course: Veterinary
Keywords: goose paramyxovirus disease monoclonal antibody colloidal gold immunochromatographic
CLC: S858.33
Type: Master's thesis
Year: 2012
Downloads: 72
Quote: 0
Read: Download Dissertation

Abstract


Goose paramyxovirus disease also known as goose newcastle disease, is a viraldisease caused by the goose source paramyxovirus designated avian paramyxovirus1with high morbidity, mortality and infection rates. ND outbreaks in geese with severeclinical signs was first reported in1997in Jiangsu province of China then was foundepidemic in many regions of the country within several years causing great economiclosses which is seriously threatenning the healthy development of geese industry.Diagnosing goose paramyxovirus disease promptly to cull infected geese in theshortest time is significant for prevention more infection and greater economic losses.Commonly methods used for goose paramyxovirus disease diagnosis includingisolation and identification of the virus, HA-HI test, agar diffusion test, ELISA,immunofluorescence, RT-PCR, real-time quantitative PCR are inconvenient to use onfield. Colloidal gold immunochromatography test strip using nitrocellulose membraneas the reaction carrier, colloidal gold as the reaction tracer and immunoadsorption asworking principle has advantages of simple to use, visible to the naked eyes, hence isconsidered suitable for rapid diagnosis of goose paramyxovirus disease on field.In this study, the NA-1strain of goose source paramyxovirus which werepurified by sucrose linear density gradient centrifugation after embryo proliferationwere used for immunization of6-8weeks’ BALB/c mice in assordance withconventional procedures. Mice with the serum titer of1:120800were chose to provideimmune spleen cells for cell fusion with SP2/0myeloma cells in the logarithmicgrowth phase. An indirect ELISA method was established for screening positivehybridoma cell lines. Three lines of hybridoma cells named3E9、2B8、1D8with thetiter of1：6400、1：3200、1:3200respectively were obtained after limited dilutioncloning more than three times.The average chromosome numbers of3E9、2B8、1D8cells was respectively98、91and87in line of the hybridoma cells. Specificity of the three monoclonal antibodies were identified by indirect ELISA with9strains of virus, including goosesource paramyxovirus NA-1、national standard virulent strain of chicken newcastledisease F48E9and chicken source newcastle disease viru strain La Sota, and otheravian disease virus, such as AIV、IBV、IBDV and MDV. All of the three monoclonalantibodies has positive reaction to strains of goose paramyxovirus NA-1、nationalstandard virulent strain of chicken newcastle disease F48E9and chicken sourcenewcastle disease viru strain La Sota, while was negative to other virus. Antibodysubclasses of3E9,2B8and1D8was IgG1， IgM and IgM respectively. Ascitesprepared by3E9hybridoma cells had a ELISA titer of1:160000. Hemagglutinationinhibition test of3E9ascites to the7strains of virus mentioned before was taken. TheHI titer of3E9to NA-1、F48E9、La Sota strains respectively was214、212、211，andno hemagglutination inhibition was found to other4strains of virus. Result of theindirect immunofluorescence assay has shown that the3E9ascites was able torecognize the NA-1、F48E9、La Sota strains specifically. The concentration of theascites after purified was2.13mg/mL, and the ELISA titer was1:128000. Polyclonalantibody anti-NA-1with the concentration of4.26mg/mL and the titer of1:640000was prepared by two adult rabbits.Immune colloidal gold strip for diagnosis of goose paramyxovirus disease wasprepared by using3E9McAb labele colloidal gold, rabbit anti-NA-1polyclonalantibody as the test line and goat anti–mouse IgG as the control line. The sensitivityof the strip was8times of the HA-HI assay for goose paramyxovirus, and2times ofthe HA-HI assay for F48E9and La Sota strains. The results of the detection to throatswabs and anal swabs from goose by this immune colloidal gold strip were88%and98%matched with the HA-HI assay of goose anti-NA-1positive serum. The immunecolloidal gold strip for diagnosis of goose paramyxovirus disease prepared in thisstudy provides a simple method for rapid diagnosis of goose paramyxovirus diseaseon field.

Related Dissertations

  1. Study on Bispecific Monoclonal Antibody to Pesticide Imidacloprid and Parathion-Methyl,S482.2
  2. Expression and Purification of Influenza a Virus M2 Protein and Detection of Its Antigenicity,R392
  3. Studies on Dynamic Regularity of Antibody Against G. Anatis and Development of Hybridoma Cell Lines Secreting Monoclonal Antibodies Against G. Anatis Lps,S858.32
  4. Development of Hybridoma Cell Lines Secreting Monoclonal Antibodies Against Lps Antigen of B. Melitensis,R392
  5. Epidemiologic Studies on Duck Circovirus Infection in East China and Development of Monoclonal Antibody Against Duck Circovirus,S858.32
  6. Purification of Recombinant Schistosoma Japonicum Protein SjGST and Preparation and Characterization of the Monoclonal Antibody,R392
  7. Cloning, Identification and Eukaryotic Expression of Variable Region of Monoclonal Antibodies Against Chelated Mercury, Copper and Zinc and Three Dimentional Modeling of Recombinant Antibody,X171.5
  8. A Research on Soil Transmission Medium of Rice Stripe Virus,S435.111.42
  9. Development of the Colloidal Gold Immuno-Chromatographyic Strips for the Early Pregnancy Diagnosis of Hu Sheep,S858.26
  10. The Study on the Preparation of McAb to Anti-Melemine and Establishment of Ic-ELISA for Detetion of Melamine,S859.84
  11. Preparation and Identification of Monoclonal Antibodies Against Bovine NRAMP1 N-Terminal Fragment,R392
  12. Genetic Study of IL-15 in All and Generation of Anti-IL-1α Antibody and Related Study of IL-1α,R733.71
  13. Establishment of Human CD48 Transgenic Cell Line and Preparation of Anti-human CD48 Monoclonal Antibody,R730.3
  14. The Experimental Study of 125I Labelled Anti-CD28 Monoclonal Antibody 2F5,R392
  15. Establishment and Application of New Detection Methods Based on gG Gene for Bovine Infectious Rhinotracheitis Virus,S858.23
  16. Establishment of Cell Line Transfected with Human GL50 Gene and Generation of Anti-human GL50 Monoclonal Antibodies with Biological Funcation,R392
  17. Construction of Human B7-H4 Transfected Cells and Preparation of Anti-human B7-H4 Monoclonal Antibody,R730.3
  18. Establishment of an Enzyme-linked Immunosorbent Assay for Measurement of Von Willebrand Factor Ristocetin Cofactor Activity and Its Applicatin in Clinic,R554.1
  19. Generation of Anti-human ppGalNAc-T2 Monoclonal Antibody and the Effect of ppGalNAc-T2 Gene Expression Mediated by Lentivirus on the Malignant Phenotype of Leukemia Cell Line,R733.7
  20. Preparation of Type HPV16 E7 Protein Monoclonal Antibody,R737.33
  21. The Expression and Fuction of IL-1β in Adult AML,R733.71

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Poultry > Goose
© 2012 www.DissertationTopic.Net  Mobile