致奶牛乳腺炎葡萄球菌快速诊断及药敏试验方法的改进

发布时间：2019-04-23 09:22

【摘要】：为了进一步提高致奶牛乳腺炎诊断方法的快速、简便和实用性,本研究在已有葡萄球菌选择培养基基础上,以刃天青钠作为指示剂,成功对致奶牛乳腺炎葡萄球菌快速诊断及药敏试验方法进行改进。改进的mBP-RS培养基具有良好的特异性和敏感性,葡萄球菌在其中生长良好,并且颜色发生明显变化,不支持大肠杆菌和链球菌等细菌生长；模拟奶样与细菌纯培养试验结果一致；能在乳腺炎相关细菌混合组合中鉴定出葡萄球菌。采集临床奶样101份,用mBP-RS培养基进行快速诊断,结果表明能在12h内对是否有葡萄球菌感染作出明确诊断,并对感染程度做出判断,试验结果既可用肉眼观察,也可用分光光度计检测。用改进mBP-RS培养基对101份奶样进行快速诊断试验,结果显示葡萄球菌单独感染31份,混合感染33份,与标准法比较,诊断敏感性可达98.3%,特异性可达95%。为了建立乳腺炎相关葡萄球菌快速药敏试验方法,在mBP-RS培养基基础上,用不同浓度抗生素进行葡萄球菌快速药敏试验,并与标准纸片扩散法比较,发现mBP-RS培养基对少数抗生素的抑菌活性有不同程度的影响,需要对其浓度进行适当校正。用校正浓度的抗生素进行快速药敏试验,所得结果与纸片法一致。对模拟奶样进行快速药敏试验,结果与细菌纯培养结果高度一致。对临床奶样进行葡萄球菌快速药敏试验,所得结果与标准方法符合率可以达到95.8%。临床奶样所含细菌对氨苄青霉素、青霉素G耐药,对氧氟沙星、头孢噻肟、头孢唑啉普遍敏感,对链霉素、红霉素、新生霉素、四环素的敏感程度因奶样不同而异。致奶牛乳腺炎葡萄球菌快速诊断及药敏试验试剂盒的改进,可以更加快速直观地对临床奶样进行检测,对有否葡萄球菌感染及其抗生素敏感性做出明确的诊断,并具有特异性强、灵敏性高等优点。特别适用于基层中小型奶牛场,拥有广阔的应用前景。
[Abstract]:In order to further improve the rapid, simple and practical diagnosis of mastitis in dairy cows, on the basis of staphylococcal selection medium, sodium azurol was used as indicator. The rapid diagnosis and drug sensitivity test of staphylococcus mastitis in dairy cows were improved successfully. The improved mBP-RS medium had good specificity and sensitivity, in which staphylococci grew well and the color changed obviously, which did not support the growth of bacteria such as Escherichia coli and Streptococcus. The simulated milk sample was consistent with the bacterial pure culture test, and staphylococcus could be identified in the mixed combination of mastitis-related bacteria. 101 clinical milk samples were collected for rapid diagnosis with mBP-RS medium. The results showed that staphylococcal infection could be diagnosed clearly within 12 hours, and the degree of infection could be judged. The results could be observed by naked eye. It can also be detected by spectrophotometer. The improved mBP-RS medium was used for rapid diagnosis of 101 milk samples. The results showed that 31 cases were infected with staphylococcus alone and 33 cases with mixed infection. Compared with the standard method, the sensitivity and specificity of staphylococcal infection were 98.3% and 95% respectively. In order to establish a rapid susceptibility test method for staphylococcus mastitis-associated staphylococcus, the rapid susceptibility test of staphylococci was carried out with different concentrations of antibiotics on the basis of mBP-RS medium, and compared with the standard disk diffusion method. It was found that the mBP-RS medium had different effects on the antibacterial activity of a few antibiotics, and the concentration of the medium needed to be properly corrected. The results of rapid drug sensitivity test with corrected concentration of antibiotics were consistent with those obtained by disk method. The results of rapid drug sensitivity test of simulated milk samples were in good agreement with the results of pure culture of bacteria. The rapid drug sensitivity test of staphylococci in clinical milk samples showed that the coincidence rate between the results and the standard method was 95.8%. The bacteria in clinical milk samples were resistant to ampicillin, penicillin G, ofloxacin, cefotaxime and cefazolin. The sensitivity to streptomycin, erythromycin, neomycin and tetracycline varied with milk samples. The rapid diagnosis of staphylococcus mastitis in dairy cows and the improvement of drug sensitivity test kit can detect the clinical milk samples more quickly and intuitively, and make a definite diagnosis of staphylococcal infection and antibiotic sensitivity. It has the advantages of strong specificity and high sensitivity. Especially suitable for grass-roots small and medium-sized dairy farms, has a broad application prospects.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.23

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