Study Of PCR Detection Method For Fowl Cholera Based On Ptfa Gene

Fowl cholera, also known as the poultry hemorrhagic septicemia or Avian Pasteurellosis, caused by Pasteurella multocida, a kind of poultry diseases. Pasteurella multocida is weaked on dug, sunlight and drying. So the disease mostly occurs in the muggy and humid environmental conditions. All kinds of poultry, such as chickens, turkeys, geese, and ducks can be infected. The disease is a very high morbidity and mortality. We treated with antibiotics when the poultry infection fowl cholera. It maybe lead to have excessive antibiotic residues in poultry meat products.Currently, most of people still judge on the basis of clinical symptoms and the separation of pathogenic bacteria to diagnosis of fowl cholera. Although the tradition method can obtion reliable results, these methods have a heavy workload and complicated operation, it also takes a long time. It is detrimental to diagnosis and control fowl cholera. It is in urgent need of a rapid, specific and sensitive diagnostic test.In this study, the genome was extracted from strains of avian Pasteurella multocida CVCC474, the ptfa gene was amplified by PCR using the specific primers which designed according to the sequence logined in GenBank. Then the reaction conditions of PCR were optimized from annealing temperature, cycle times, concentrion of primers, concentration of Mg2+and concentration of dNTP, accompanied with the studying on the specificity and sensitivity of PCR. The results showed that the435bp DNA fragment was specifically amplified from avian Pasteurella multocida and no DNA fragment was obtained from other pathogenic bacteria including chicken pathogenic Escherichia coli, Chicken Staphylococcus aureus and Salmonella pullorum. The sensitivity test showed that the sensitivity of this PCR method was1pg/μL. In conclusion, this experiment developed the foundation for the research of rapid detection method against avian Pasteurella multocida infection, and made some contribution to the security of our country about the poultry products.