本文選題：聚合酶鏈?zhǔn)椒磻?yīng) + 氣腫疽梭菌�。� 參考：《延邊大學(xué)》2015年碩士論文

【摘要】：氣腫疽是由氣腫疽梭菌引起的以牛為主的一種熱性、急性、地方性傳染病,以肌肉和深部組織發(fā)生氣性壞疽為主要特征。該病主要經(jīng)過(guò)皮膚及消化道創(chuàng)傷感染,往往來(lái)不及治療就會(huì)導(dǎo)致病畜死亡,羊、豬、鹿甚至是人也都有相關(guān)報(bào)道顯示被感染,被國(guó)際獸醫(yī)局評(píng)定為第三類傳染病。氣腫疽在0.5～3歲的黃牛及水牛多發(fā),無(wú)明顯的季節(jié)規(guī)律。氣腫疽在我國(guó)的暴發(fā),給我國(guó)的畜牧業(yè)帶來(lái)了巨大的經(jīng)濟(jì)損失,也引起了人們的高度關(guān)注。但目前尚無(wú)有效的防控手段,因此,建立快速、敏感且特異的檢測(cè)方法來(lái)診斷氣腫疽病是當(dāng)務(wù)之急。本試驗(yàn)為建立氣腫疽梭菌(Clostridium chauvoei)的快速檢測(cè)方法,根據(jù)GenBank中已發(fā)表的氣腫疽梭菌細(xì)胞毒素cctA基因序列(登錄號(hào)：JQ692583)設(shè)計(jì)了一對(duì)特異性引物,并以氣腫疽梭菌標(biāo)準(zhǔn)株C54-1全基因組DNA為模板,對(duì)氣腫疽梭菌cctA基因進(jìn)行PCR擴(kuò)增,并經(jīng)過(guò)敏感性和特異性試驗(yàn)對(duì)該試驗(yàn)方法進(jìn)行驗(yàn)證。結(jié)果顯示：建立的氣腫疽梭菌PCR檢測(cè)方法擴(kuò)增出的片段大小為501bp,與GenBank上氣腫疽梭菌的同源性達(dá)99.4%,且該方法與D型產(chǎn)氣莢膜梭菌、E型產(chǎn)氣莢膜梭菌、腐敗梭菌和巴氏桿菌等病原體無(wú)交叉反應(yīng),最低檢測(cè)濃度為12.30 fg/μL。結(jié)果表明,建立的PCR檢測(cè)方法具有快速、特異、敏感、高效等優(yōu)點(diǎn),與其他診斷方法相比更適用于氣腫疽梭菌的診斷,為牛氣腫疽病的快速檢測(cè)和診斷提供基礎(chǔ)研究。
[Abstract]:Emphysema is a hot, acute, endemic infectious disease caused by Clostridium emphysema. It is characterized by the occurrence of gaseous gangrene in muscles and deep tissues.The disease mainly passes through the skin and the digestive tract wound infection, often causes the disease animal death without the time treatment, sheep, the pig, the deer and even the human also has the related report to show to be infected, is rated as the third kind infectious disease by the international veterinary bureau.Emphysema was more common in 3-year-old yellow cattle and buffalo with no obvious seasonal regularity.The outbreak of emphysema in our country has brought huge economic loss to animal husbandry in our country.However, there are no effective methods to prevent and control emphysema, so it is urgent to establish a rapid, sensitive and specific detection method for the diagnosis of emphysema.In order to establish a rapid detection method for Clostridium chauvoeii, a pair of specific primers were designed according to the cctA gene sequence of Clostridium chauvoeii published in GenBank (accession number: JQ692583).The whole genome DNA of C. emphysema strain C54-1 was used as a template to amplify the cctA gene of Clostridium emphysema by PCR, and the method was verified by sensitivity and specificity test.The results showed that the amplified fragment size of the established PCR detection method was 501bp, and the homology was 99.4 with that of Clostridium emphysema on GenBank, and the method was similar to that of Clostridium perfringens type D (Clostridium perfringens).There was no cross reaction between Clostridium putrefaciens and Pasteurella, and the lowest detection concentration was 12.30 fg/ 渭 L.The results showed that the established PCR detection method was rapid, specific, sensitive and efficient. Compared with other diagnostic methods, it was more suitable for the diagnosis of Clostridium emphysema, and provided the basic research for rapid detection and diagnosis of bovine emphysema.
【學(xué)位授予單位】：延邊大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S858.23

【參考文獻(xiàn)】

相關(guān)期刊論文 前8條

1 陳瑩瑩;金鑫;姜成國(guó);婁安鋼;姜丹丹;汪雪燁;鄧文學(xué);李香春;;氣腫疽梭菌菌體抗原分析[J];安徽農(nóng)業(yè)科學(xué);2012年17期

2 郭洪友;鄭聃;;牛氣腫疽病的診治[J];現(xiàn)代農(nóng)業(yè)科技;2010年07期

3 鄭景生,呂蓓;PCR技術(shù)及實(shí)用方法[J];分子植物育種;2003年03期

4 孫瑛;雷國(guó)云;于林潔;;牛氣腫疽診斷與綜合防治[J];中國(guó)畜禽種業(yè);2008年17期

5 關(guān)孚石,張立春,張瑞婷,田振詳,葛展義,張志軍;牛氣腫疽病的診斷及防治[J];中國(guó)動(dòng)物檢疫;1998年05期

6 于桂陽(yáng),陳松明,黃杰河;黃牛氣腫疽的診治[J];中國(guó)獸醫(yī)科技;2002年03期

7 楊利;劉良;曹三杰;文心田;黃小波;;豬胸膜肺炎放線桿菌半套式PCR檢測(cè)方法的建立及應(yīng)用[J];中國(guó)獸醫(yī)科學(xué);2009年09期

8 王光明;李萬(wàn)財(cái);萬(wàn)么才郎;鄂艷花;諾科加;才旦卓瑪;;藏系綿羊氣腫疽的診治[J];中國(guó)獸醫(yī)雜志;2008年05期

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