【摘要】：為了進一步提高致奶牛乳腺炎診斷方法的快速、簡便和實用性,本研究在已有葡萄球菌選擇培養(yǎng)基基礎(chǔ)上,以刃天青鈉作為指示劑,成功對致奶牛乳腺炎葡萄球菌快速診斷及藥敏試驗方法進行改進。改進的mBP-RS培養(yǎng)基具有良好的特異性和敏感性,葡萄球菌在其中生長良好,并且顏色發(fā)生明顯變化,不支持大腸桿菌和鏈球菌等細菌生長；模擬奶樣與細菌純培養(yǎng)試驗結(jié)果一致；能在乳腺炎相關(guān)細菌混合組合中鑒定出葡萄球菌。采集臨床奶樣101份,用mBP-RS培養(yǎng)基進行快速診斷,結(jié)果表明能在12h內(nèi)對是否有葡萄球菌感染作出明確診斷,并對感染程度做出判斷,試驗結(jié)果既可用肉眼觀察,也可用分光光度計檢測。用改進mBP-RS培養(yǎng)基對101份奶樣進行快速診斷試驗,結(jié)果顯示葡萄球菌單獨感染31份,混合感染33份,與標準法比較,診斷敏感性可達98.3%,特異性可達95%。為了建立乳腺炎相關(guān)葡萄球菌快速藥敏試驗方法,在mBP-RS培養(yǎng)基基礎(chǔ)上,用不同濃度抗生素進行葡萄球菌快速藥敏試驗,并與標準紙片擴散法比較,發(fā)現(xiàn)mBP-RS培養(yǎng)基對少數(shù)抗生素的抑菌活性有不同程度的影響,需要對其濃度進行適當校正。用校正濃度的抗生素進行快速藥敏試驗,所得結(jié)果與紙片法一致。對模擬奶樣進行快速藥敏試驗,結(jié)果與細菌純培養(yǎng)結(jié)果高度一致。對臨床奶樣進行葡萄球菌快速藥敏試驗,所得結(jié)果與標準方法符合率可以達到95.8%。臨床奶樣所含細菌對氨芐青霉素、青霉素G耐藥,對氧氟沙星、頭孢噻肟、頭孢唑啉普遍敏感,對鏈霉素、紅霉素、新生霉素、四環(huán)素的敏感程度因奶樣不同而異。致奶牛乳腺炎葡萄球菌快速診斷及藥敏試驗試劑盒的改進,可以更加快速直觀地對臨床奶樣進行檢測,對有否葡萄球菌感染及其抗生素敏感性做出明確的診斷,并具有特異性強、靈敏性高等優(yōu)點。特別適用于基層中小型奶牛場,擁有廣闊的應(yīng)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.
【學位授予單位】：揚州大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S858.23

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