摘要:
应用血清抗体间接ELISA方法对福建省8个地市的23个猪场采集的378份血清进行Hps抗体检测;采集疑似副猪嗜血杆菌感染病例的病料,用脱纤马血TSA平板对Hps进行分离与鉴定;用琼扩试验对分离株进行血清型鉴定;根据GenBank登录的HS0165株的ompP5基因序列合成1对特异性引物,提取分离菌株的基因组DNA进行PCR扩增,回收目的片断并进行克隆、测序和同源性分析。结果显示,福建省23个猪场的Hps场阳性率为100%,猪群阳性率为36.78%;从疑似感染Hps的猪只中共分离到52株Hps,分别为血清2型、4型、5型、13型和未定型,其中血清13型Hps分离最多; 3株Hps 分离株的外膜蛋白P5基因(ompP5)序列与GenBank中提交序列的同源性达到了93%以上。
Abstract:
Hps antibody of 378 serum sample from 23 pig farm in 8 city of Fujian Province was detected by the indirect ELISA reagent kit; And Hps was separated and identified by use of TSA plate with removal of fine horse blood from the collected disease materials of swine suspected infecting Hps; And the serotype of separated strains was classified by the Agar diffusion test; And a pair of specific primers was synthesized by referring to ompP5 gene of HS0165 strain signed in GenBank, and the ompP5 gene was amplified using Hps DNA extracted as a template by PCR method, cloned, sequenced and blasted. The results showed that the positive rate in 23 field from Fujian Province was to 100%, and the positive rate of the herd was to 36.78%; 52 isolates from the swine of suspected infecting Hps had been separated and identified, and were regarded as serotype 2, 4, 5, 13 and non-serotype respectively, in addition, the serotype 13 was the most; Compared the GenBank, the homology of ompP5 of 3 isolates are respectively up to 93%.
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