本文選題：諾卡氏菌屬 + SecA基因��； 參考：《中國人獸共患病學(xué)報》2017年06期

【摘要】：目的將諾卡氏菌屬細(xì)菌的一段特異性DNA設(shè)計成分子信標(biāo)探針,用于該細(xì)菌的PCR檢測。方法將諾卡氏菌屬細(xì)菌、戈登氏菌屬細(xì)菌及紅球菌屬細(xì)菌菌株分別接種于腦心浸液瓊脂培養(yǎng)基分離培養(yǎng),觀察其生長情況,提取菌株DNA作為擴(kuò)增模板;設(shè)計諾卡氏菌屬細(xì)菌基于secA1基因的特異性分子信標(biāo)探針,在實時熒光定量PCR反應(yīng)譯體系中加入分子信標(biāo)探針,PCR產(chǎn)物進(jìn)行熒光信號檢測。結(jié)果諾卡氏菌secA1基因經(jīng)實時熒光定量PCR擴(kuò)增后可產(chǎn)生陽性熒光信號,紅球菌屬細(xì)菌及戈登氏菌屬細(xì)菌的secA1基因、陰性對照實驗組及空白對照組經(jīng)實時熒光定量PCR擴(kuò)增后不產(chǎn)生熒光信號,為陰性。結(jié)論 secA1作為看家基因,是用來進(jìn)行種水平的鑒定及系統(tǒng)進(jìn)化研究非常理想的靶分子,而分子信標(biāo)探針技術(shù)可以準(zhǔn)確、快速、靈敏的進(jìn)行諾卡氏菌secA1基因檢測。
[Abstract]:Objective to design a molecular beacon probe for the detection of Nocardia bacteria by using a segment of specific DNA. Methods the strains of Nocardia, Gordon's and Rhodococcus were isolated and cultured on the Agar medium of brain heart extract, and their growth was observed. The strain DNA was extracted as a template for amplification. A specific molecular beacon probe based on secA1 gene was designed for Nocardia bacteria. The fluorescent signal was detected by adding a molecular beacon probe into the system of real-time fluorescence quantitative PCR reaction. Results the secA1 gene of Nocardia spp. was amplified by real-time fluorescence quantitative PCR, and the secA1 gene of Rhodococcus and Gordenia spp. The negative control group and the blank control group did not produce fluorescence signal after real-time fluorescence quantitative PCR amplification. Conclusion as a housekeeping gene, secA1 is an ideal target molecule for species identification and phylogenetic study, and molecular beacon probe technique can be used to detect the secA1 gene of Nocardial bacillus accurately, quickly and sensitively.
【作者單位】： 貴州醫(yī)科大學(xué)微生物學(xué)教研室;貴州醫(yī)科大學(xué)生化與分子生物學(xué)教研室;
【基金】：國家自然基金(No.31260029) 貴州省國際科技合作項目(黔科合外G字[2014]7006) 貴陽市人民政府-貴州醫(yī)科大學(xué)科學(xué)技術(shù)聯(lián)合基金“貴陽市科技局創(chuàng)新團(tuán)隊”項目(GY2015-18) 貴州省科技計劃項目(黔科合[2010]3154號)聯(lián)合資助~~
【分類號】：R440
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本文編號：1849969