Dissertation > Excellent graduate degree dissertation topics show

Cloning and Expression of CP15/60 and CP41 Gene of Cryptosporidium Parvum Mouse Genotype and Epidemiology Survey of Pig Cryptosporidiosis in One Hoggery in Shanghai District

Author: YuHuiZhu
Tutor: YueCheng;ChenZhaoGuo
School: Xinjiang Agricultural University
Course: Preventive Veterinary Medicine
Keywords: Cryptosporidium CP15/60 and CP41 cloning and expression ELISA epidemiology survey Pig
CLC: S852.723
Type: Master's thesis
Year: 2008
Downloads: 75
Quote: 0
Read: Download Dissertation


Cryptosporidium is a world-wide waterborne protozoan zoonosis and harm severely to humans and animals. It grows and reproduces within epithelial cells of the digestive and respiratory organs of verterbrates with a wide host range, including humans and animals belong to mammals, birds, reptiles and fish. Cryptosporidium infection can lead to persistant, severe and often life-threatening disease in immunocompromized patients and animals. To date, there is still no effective treatment available for cryptosporidiosis in man and animals. It’s necessary to develop sensitive and rapid diagnosis technique and effective preventative vaccine.The sporozoite surface antigen CP15/60 and the oocyst wall protein CP41 genes were successful amplified from total RNA of Cryptosporidium parvum mouse genotype by reverse transcription polymerase chain reaction(RT-PCR). The homology of the nucleotide sequences between cloned genes and the genes of C. parvum bovine genotype downloaded from GenBank were 98.66 % and 90.50 %, respectively.The prokaryotic expressed plasmid pET-28b(+)-CP15 and pGEX-5x-3-CP41 were constructed and expressed in E. coli successfully. Western-blot analysis showed that the recombinant expressed proteins could be recognized by the sera of rabbit infected with C. parvum. ELISA results showed that the titer of specific antibody against the recombinant proteins in sera from 3 times immunized SPF rabbits were significantly higher than that of rabbits before immunization, indicating that the recombinant proteins have good immune-reactivity and immunogenicity.Indirect ELISA techniques were developed for detecting specific antibody against Cryptosporidium with the recombinant protein CP15/60 and CP41 as the coating antigen respectively. The techniques were used to detect the specific antibody titre of sera of rabbits infected with C. parvum rabbit genotype or not. The results indicated that the indirect ELISA technique has high sensitivity and specificity.Surveying systematically and extensively of epidemiology of cryptosporidiosis is a basic work for preventing the disease effectively. Infection of Cryptosporidium in pig was surveyed in one hoggery in Shanghai district. 1,629 samples were collected and detected by Sheather’s sucrose floatation technique. Results showed that the infection rate of Cryptosporidium in pig was 31.55 %. In one year time, spring and winter are two seasons with the highest infection rate, and then the autumn. The summer has the lowest infection rate. The infection rate also related with the ages of pigs. The pigs at 30-39 and 40-49 days old have the highest infection rate. They were 58.54 % and 43.30 %, respectively. In contrast, the infection rate is very low when pigs smaller than 30-day-old or exceed 90-day-old. The infection intensity of Cryptosporidium in pigs also related with their ages, younger pigs has higher infection intensity.The work was much helpful to know the infection condition about Cryptosporidium in pigs in Shanghai district and to control the disease. The cloning and expression of sporozoite surface antigen CP15/60 and oocyst wall protein CP41 gene and the development of indirect ELISA diagnosis technique for cryptosporidiosis would also be much helpful for next serology survey of the disease and researching of immunological prevention technique with the two recombinant proteins.

Related Dissertations

  1. Study on Bispecific Monoclonal Antibody to Pesticide Imidacloprid and Parathion-Methyl,S482.2
  2. cDNA Cloning, Expression of vp5 and vp7 Genes and Subcecullar Localization of VP5 and VP7 Proteins in Grass Carp Reovirus,S941.41
  3. Cloning, Expression of vp6 and ns38 Genes and Immunogenicity of VP6 and NS38 in Grass Carp Reovirus,S941.41
  4. Research of the Correlation About the Expression of Mfap4 in Liver Fibrosis Tissues and Concentrations of Mfap4 in Peripheral Blood Associated with Liver Pathological Grade,R575.2
  5. Establishment of ELISA for Detecting Cd-MT of Pteria Penguin and Preliminary Study of ELISA-kit,X835
  6. Characterization and Identification of Maturity of Pig Manure Composting,S141.4
  7. Polymorphisms in the Promoter Region of Swine BMP7 Gene and Their Association with Reproductive Traits,S828
  8. Establishment of Inderect ELISA for Detection of Antibodies Angainst STLV-1 and Preparation of Hybridoma Cell of Monoclonal Antibodies,R373
  9. Study on the Combined Probiotics and Its Effects on Prodution Performance of Pigs,S828.5
  10. Studies on Dynamic Regularity of Antibody Against G. Anatis and Development of Hybridoma Cell Lines Secreting Monoclonal Antibodies Against G. Anatis Lps,S858.32
  11. Purification of Human Respiratory Syncytial Virus Fusion Protein with Immunomagnetic Microsphere Technology,R373
  12. Isolation and Characterization of Carbendazim-Degrading Strains, Cloning and Expression of the mheI Gene,X172
  13. Development of Hybridoma Cell Lines Secreting Monoclonal Antibodies Against Lps Antigen of B. Melitensis,R392
  14. Construction and Application of the Integrated Gene Containing Multi-Mimotopes and VP2 of Infectious Bursal Disease Virus,S852.65
  15. The Expression of CPV-2 VP2 Gene in SF9 and Development of Indirect Elisa for Serum Antibodys,S852.65
  16. Pathogenic Mechanism of Invasion Associated Genes in Avian Pathogenic E. Coli Duck Isolate DE205B,S852.61
  17. Isolation, Identification and Degrading Characteristic of a Cyhalofop-Butyl-Degrading Bacteria, and Cloning, Expression of a Cyhalofop-Butyl-Hydrolyzing Gene chbH,X172
  18. Cloning and Expression of Organic Solvent Tolerant Lipase Gene from Staphylococcus Saprophyticus M36,Q78
  19. Screening of Bacterial Strains with High α-Galactosidase Activity and Its Gene Cloning, Expression, Purification and Characterization,TQ925
  20. The Research on Enzyme-Linked Immunosorbent Assay for Pretilachlor and Acetochlor,S482.4
  21. Production of Monoclonal Antibodies Against Thiabendazole and Development of Heterologos Elisa,S482.2

CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Livestock parasitology > Veterinary protozoan school > Cryptosporidium
© 2012 www.DissertationTopic.Net  Mobile