本文選題：豬種布魯菌 + 牛種布魯菌　； 參考：《西北農(nóng)業(yè)學(xué)報(bào)》2017年08期

【摘要】：為建立在臨床樣本能夠同時(shí)檢測(cè)豬種、牛種、羊種、綿羊種布魯菌的多重PCR方法,根據(jù)NCBI已收錄的布魯菌全基因,設(shè)計(jì)并合成4對(duì)特異性引物,通過優(yōu)化多重PCR的反應(yīng)條件,建立能夠同時(shí)檢測(cè)4種布魯菌的多重PCR診斷方法。特異性試驗(yàn)結(jié)果表明,可以從4種布魯菌以及4種細(xì)菌混合物中擴(kuò)增出大小分別為276、494、733和976bp的特異性條帶,對(duì)照組的檢測(cè)結(jié)果為陰性;敏感性試驗(yàn)結(jié)果表明,對(duì)4種病原菌基因組DNA的檢出量為豬種32.2pg、牛種21.3pg、羊種27.5pg、綿羊種43.2pg;人工模擬感染樣本檢測(cè)結(jié)果表明,能從混合感染的病料中特異地檢測(cè)出4種病原菌。應(yīng)用該方法對(duì)200份臨床奶山羊乳樣進(jìn)行檢測(cè),結(jié)果檢出4份陽性。建立的多重PCR方法具有特異性強(qiáng)、敏感度高、穩(wěn)定性好的特點(diǎn),可以有效地檢測(cè)4種布魯菌的感染。
[Abstract]:In order to establish a multiplex PCR method for simultaneous detection of brucellosis in swine, cattle, sheep and sheep in clinical samples, four pairs of specific primers were designed and synthesized according to the whole genes of Brucella collected by NCBI, and the reaction conditions of multiplex PCR were optimized. A multiplex PCR method for simultaneous detection of four Brucella species was established. The specific bands of 276494733 and 976bp could be amplified from four Brucella species and four bacterial mixtures, respectively. The results of the control group were negative, and the sensitivity test showed that the specific bands could be amplified from the four Brucella species and the four bacterial mixtures. The amount of genomic DNA of four pathogens was 32.2pg for pigs, 21.3pg for cattle, 27.5pg for sheep and 43.2pg for sheep. The method was used to detect 200 milk samples of clinical dairy goats and 4 samples were positive. The multiplex PCR method has the characteristics of high specificity, high sensitivity and good stability. It can be used to detect the infection of four Brucella species effectively.
【作者單位】： 西北農(nóng)林科技大學(xué)動(dòng)物醫(yī)學(xué)院;
【基金】：陜西省農(nóng)業(yè)科技創(chuàng)新與攻關(guān)(2016NY-092) 陜西省重點(diǎn)產(chǎn)業(yè)創(chuàng)新鏈(2016KTZDNY02-06) 西北農(nóng)林科技大學(xué)試驗(yàn)示范站(基地)科技成果推廣(TGZX2015-32)~~
【分類號(hào)】：S852.61

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 許鄒亮;南文龍;陳義平;;分子生物學(xué)技術(shù)在布魯菌種型鑒定上的應(yīng)用[J];動(dòng)物醫(yī)學(xué)進(jìn)展;2012年01期

2 楊艷玲;唐婕;白光彥;盛雪玲;王成福;王興龍;;羊布魯菌膜蛋白的提取及血清抗體間接ELISA檢測(cè)方法的建立[J];中國生物制品學(xué)雜志;2010年04期

3 吳小艷;王希良;;布魯菌與巨噬細(xì)胞相互作用的研究進(jìn)展[J];微生物學(xué)免疫學(xué)進(jìn)展;2008年01期

4 李秀梅;郭戀;梁智選;李穎;郭立力;石瑜;朱雅寧;王紅軍;;布魯菌環(huán)介導(dǎo)等溫?cái)U(kuò)增檢測(cè)試劑盒的研制[J];動(dòng)物醫(yī)學(xué)進(jìn)展;2014年07期

5 駱利敏;崔步云;趙鴻雁;芮勇宇;李明;;脈沖場(chǎng)電泳分析布魯菌標(biāo)準(zhǔn)菌株[J];檢驗(yàn)醫(yī)學(xué);2006年05期

6 鄧麗芳;黃亞寧;羅雯;;布魯菌病疫苗研制進(jìn)展[J];陜西農(nóng)業(yè)科學(xué);2012年02期

7 趙忠鵬;王希良;;布魯菌毒力因子研究進(jìn)展[J];微生物學(xué)免疫學(xué)進(jìn)展;2007年01期

8 徐杰;于爽;付思美;鐘志軍;王玉飛;曲R，

本文編號(hào)：2032935