关键词: 盘羊, PCR, 绵羊肺炎支原体

Abstract: To detect infection of Mycoplasma ovipneumoniae (MO)of Argali (Ovis ammon),we designed a pair of specific primers to amplify a 406 bp fragment of the 16S rRNA gene of MO. After analysis of the optimization reaction condition, e. g. ,the specificity and sensitivity for PCR of the MO gene,anneal temperature,concentration of Taq DNA polymerase and that of the primers,the sensitivity of PCR was found to be 0.4075 ng genome DNA for MO. A 406 bp fragment was amplified from standard strain of MO Y98,and another was an isolated strain of MO from the secretion of ill argali. To confirm specificity,strains of Mycoplasma mycoides subsp. Capri (MMC),E. coli (EC)and Staphylococcus aureus (SA),were not able to be detected by this method. These results indicate that this test might be used to identify MO without culture or isolation. In this study,34 nasal secretions were tested by PCR and a culture-dependent method respectively.
MO was isolated from specimens of four of the animals,and 11 specimens were PCR positive. Similarly,31 serum samples of the animals were tested by antibody detection kit,and 10 seropositive animals detected. The positive rate of the three methods was 32.35% ,11.76% and 32. 26% respectively. These results show that compared to conventional methods and with related kit,the PCR method has significant economical and practical advantages. The PCR test permits ready identification of MO for diagnostic and epidemiologic study. It indicates that the PCR method is a powerful tool for identification of MO of Argali (Ovis ammon).

Key words: Argali (Ovis ammon), Mycoplasma ovipneumoniae (MO), PCR