盘羊肺炎支原体感染PCR检测方法的建立与初步应用.PDFVIP

兽类学报,2011,31 (3):312 - 316 Acta Theriologica Sinica 盘羊肺炎支原体感染 PCR 检测方法的建立与初步应用 冷青文 李志远 屈勇刚 鲁海富 金云云 王静梅 剡根强 (石河子大学动物科技学院,石河子 832003) 关键词:盘羊;PCR ;绵羊肺炎支原体 中图分类号:S85 文献标识码:A 文章编号:1000 - 1050 (2011 )03 - 0312 - 05 Establishment and application of PCR for diagnose Mycoplasma ovipneu- moniae of Argali (Ovis ammon) LENG Qingwen,LI Zhiyuan,QU Yonggang,LU Haifu,JIN Yunyun,WANG Jingmei,YAN Gen-  qiang (College of Animal Science and Technology ,Shihezi University,Shihezi 832003,China) Abstract :To detect infection of Mycoplasma ovipneumoniae (MO)of Argali (Ovis ammon),we designed a pair of spe- cific primers to amplify a 406 bp fragment of the 16S rRNA gene of MO. After analysis of the optimization reaction condi- tion,e. g. ,the specificity and sensitivity for PCR of the MO gene,anneal temperature,concentration of Taq DNA poly- merase and that of the primers,the sensitivity of PCR was found to be0 4075 ng genome DNA for MO. A406 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 with- out 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. Thes