發(fā)布時間：2018-06-15 18:01

  本文選題：水牛乳房炎 + 分離鑒定��； 參考：《廣西大學(xué)》2015年碩士論文

【摘要】：本研究對廣西奶水牛88份乳樣進行病原菌分離鑒定,根據(jù)引起奶水牛乳房炎的主要致病菌種類建立多重PCR快速診斷方法,并對該檢測方法進行優(yōu)化和臨床檢測評價。實驗內(nèi)容:1、通過對88份奶樣進行細菌分離鑒定,從56份奶樣中分離出112株病原菌,包括葡萄球菌47株,占41.9%;鏈球菌25株,占22.3%;腸桿菌21株,占18.8%;真菌12株,占10.8%;其它細菌7株,占6.2%。2、根據(jù)引起水牛乳房炎主要致病菌金黃色葡萄球菌nuc基因、大腸桿菌rrnB基因,無乳鏈球菌ef-tu基因,表皮葡萄球菌se2313基因序列設(shè)計選擇4對特異性引物,建立并優(yōu)化同時檢測這4種病原菌的4重PCR快速診斷方法。實驗結(jié)果表明:1、引起水牛乳房炎的主要致病菌為金黃色葡萄球菌、表皮葡萄球菌、無乳鏈球菌和大腸桿菌,還有部分真菌感染;感染主要以混合感染為主,56份感染奶樣中有40個為2種及以上的多種致病菌混合感染,占71.4%。2、PCR產(chǎn)物進行電泳,大腸桿菌、金黃色葡萄球菌、表皮葡萄球菌、無乳鏈球菌分別在722bp、279bp、581bp和197bp處擴增出相應(yīng)目的條帶,且無其它非特異性產(chǎn)物,實驗與臨床檢測結(jié)果證明該方法敏感性好,特異性高,與傳統(tǒng)細菌分離檢測方法相比,既保證結(jié)果準確,又大幅度縮短檢測時間,提高檢測效率。
[Abstract]:In this study, 88 milk samples of Guangxi milk buffalo were isolated and identified. The rapid multiplex PCR method was established according to the main pathogenic bacteria causing mastitis of milk buffalo, and the detection method was optimized and evaluated clinically. Experiment content: 1, through the isolation and identification of bacteria from 88 milk samples, 112 strains of pathogenic bacteria were isolated from 56 milk samples, including 47 strains of staphylococcus, accounting for 41.9%; 25 strains of streptococcus, 22.3x; 21 strains of Enterobacter, 18.8%; 12 strains of fungi, 10.8%; and 7 strains of other bacteria, According to the sequence of Staphylococcus aureus nuc gene, Escherichia coli rrnB gene, ef-tu gene and se2313 gene of Staphylococcus epidermidis, 4 pairs of specific primers were designed. To establish and optimize a 4-fold PCR rapid diagnostic method for simultaneous detection of these four pathogens. The results showed that the main pathogens causing buffalo mastitis were Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus lactococcus and Escherichia coli, and some fungal infections. 40 of 56 milk samples were mixed with two or more kinds of pathogenic bacteria, accounting for 71.4% of PCR products, E. coli, Staphylococcus aureus, Staphylococcus epidermidis, E. coli, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Staphylococcus aureus and Staphylococcus epidermidis. The corresponding target bands were amplified from 722bpn279bp581 BP and 197bp, respectively, and there were no other nonspecific products. The experimental and clinical results showed that this method was more sensitive and specific than the traditional bacterial isolation and detection methods. It not only ensures the accuracy of the results, but also greatly shortens the detection time and improves the detection efficiency.
【學(xué)位授予單位】：廣西大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S858.23

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