关键词: 鸭疫里默氏杆菌, 16SrRNA, PCR

Abstract: Primers are designed based on the conserved region of the 16SrRNA nucleotide sequences of Riemerella anatipestifer(RA) in GenBank, and the specific PCR assay for RA detection is developed. Using this assay, we can amplify a differential fragment about 844 bp from RA, but we cannot get the same fragment from Escherichia coli, Salmonella, Staphylococcus aureus, Bifidobucterium pullorum, Lactobacillus aviarius, Enterococcus faecium and Bacillus subtilis. The DNA band was obtained from agarose gel, and the sequence of the PCR product is aligned with the available sequence in GenBank. A series of sensitivity experiments is performed and proves that the detection limit of this assay is 22 pg genomic DNA and 80 CFU/m L. The clinical samples and bacteria isolated from the infected ducks are detected by the PCR assay and the positive coincidence rate comparing RA isolation with PCR is 100 percent. The result shows that the PCR assay is rapid, sensitive and specific which provides a greatly improved tool for diagnosing the infected ducks, doing molecular epidemiology survey of RA and quickly detecting the isolated bacteria.

Key words: Riemerella anatipestifer, 16SrRNA, PCR