本文選題：產(chǎn)氣莢膜梭菌　切入點(diǎn)：毒素基因　出處：《動(dòng)物醫(yī)學(xué)進(jìn)展》2017年03期

【摘要】：建立一種快速鑒別診斷不同型產(chǎn)氣莢膜梭菌的PCR檢測方法,為動(dòng)物產(chǎn)氣莢膜梭菌病的快速診斷及流行病學(xué)調(diào)查提供有效的技術(shù)手段�？朔䝼鹘y(tǒng)鑒定方法耗時(shí)長、費(fèi)用高的缺點(diǎn),提高了檢測效率。通過對產(chǎn)氣莢膜梭菌α毒素、β毒素、ε毒素和ι毒素基因序列分析,利用Premier5.0軟件設(shè)計(jì)并合成了5對特異性引物,建立了針對5種不同型產(chǎn)氣莢膜梭菌的PCR鑒別診斷方法。通過反復(fù)試驗(yàn)確定了最佳退火溫度為53℃。通過靈敏度試驗(yàn)表明,PCR檢測方法最低能檢測到的DNA濃度α毒素為308pg/μL,β毒素、ε毒素為30.8pg/μL,ι毒素A為0.122pg/μL,ι毒素B為0.05pg/μL。通過特異性試驗(yàn)表明,本方法具有較高的特異性。同時(shí),通過對本方法檢測出的陽性樣品16S rRNA序列分析發(fā)現(xiàn),與GenBank中的其他產(chǎn)氣莢膜梭菌的16S rRNA序列同源性均在98%以上。表明建立的檢測方法靈敏度高、特異性強(qiáng),可以應(yīng)用于動(dòng)物產(chǎn)氣莢膜梭菌病的實(shí)驗(yàn)室診斷。
[Abstract]:A rapid PCR detection method for the differential diagnosis of different Clostridium perfringens was established to provide an effective technique for the rapid diagnosis and epidemiological investigation of Clostridium perfringens in animals, and to overcome the disadvantages of long time and high cost in traditional identification methods. The detection efficiency was improved. By sequence analysis of 偽 toxin, 尾 toxin, 蔚 toxin and l toxin genes of Clostridium perfringens, 5 pairs of specific primers were designed and synthesized by Premier5.0 software. A method for the differential diagnosis of 5 different Clostridium perfringens by PCR was established. The optimum annealing temperature was determined to be 53 鈩，

本文編號：1661134