天堂国产午夜亚洲专区-少妇人妻综合久久蜜臀-国产成人户外露出视频在线-国产91传媒一区二区三区

水中產(chǎn)氣莢膜梭菌的熒光定量PCR檢測(cè)法

發(fā)布時(shí)間:2018-07-26 06:18
【摘要】:目的研究水中產(chǎn)氣莢膜梭菌的熒光定量PCR檢測(cè)方法,縮短水中產(chǎn)氣莢膜梭菌的檢測(cè)時(shí)間。方法根據(jù)產(chǎn)氣莢膜梭菌的α-toxin基因設(shè)計(jì)引物并擴(kuò)增,構(gòu)建α-toxin-T重組質(zhì)粒,對(duì)重組質(zhì)粒進(jìn)行梯度稀釋后進(jìn)行熒光定量PCR檢測(cè);用該方法檢測(cè)水中常見的15種細(xì)菌,確定方法的特異性;檢測(cè)產(chǎn)氣莢膜梭菌加標(biāo)水樣,驗(yàn)證方法的可行性。結(jié)果成功擴(kuò)增出產(chǎn)氣莢膜梭菌的α-toxin基因并連接T載體(417 bp)。α-toxin-T重組質(zhì)粒的稀釋度在2.17×10~0~2.17×10~9 copies/μl的范圍內(nèi),PCR反應(yīng)的特異性良好,且重組質(zhì)粒各濃度梯度間間隔的Ct值相近;所得回歸方程為y=31.69-3.11x,r=0.997 73;該方法的檢出限可達(dá)到為10 copies/μl。15株受試細(xì)菌DNA的檢測(cè)結(jié)果均為陰性,產(chǎn)氣莢膜梭菌加標(biāo)水樣檢測(cè)結(jié)果陽性。結(jié)論該方法操作快速,具有較好的特異性和靈敏性,適用于水中產(chǎn)氣莢膜梭菌的快速檢測(cè)。
[Abstract]:Objective to study the fluorescence quantitative PCR method for the detection of Clostridium perfringens in water, and to shorten the detection time of Clostridium perfringens in water. Methods according to the 偽 -toxin gene of Clostridium perfringens, primers were designed and amplified to construct 偽 -toxin-T recombinant plasmid. The recombinant plasmid was detected by fluorescence quantitative PCR after gradient dilution, and 15 common bacteria in water were detected by this method, and the specificity of the method was determined. Detection of Clostridium perfringens plus standard water samples to verify the feasibility of the method. Results the 偽 -toxin gene of Clostridium perfringens was successfully amplified and ligated into T vector. The dilution of the recombinant plasmid was 2.17 脳 10 ~ (10) ~ 0 ~ 2.17 脳 10 ~ (9) copies/ 渭 l. The regression equation was obtained as YYI 31.69-3.11xrnr 0.99773.The detection limit of this method was 10 copies/ 渭 l.15 strains of bacteria DNA were negative, and the results of Clostridium perfringens plus standard water samples were positive. Conclusion the method is rapid, specific and sensitive, and can be used for rapid detection of Clostridium perfringens in water.
【作者單位】: 中國疾病預(yù)防控制中心環(huán)境與健康相關(guān)產(chǎn)品安全所;
【基金】:國家衛(wèi)生和計(jì)劃生育委員會(huì)公益性衛(wèi)生行業(yè)科研專項(xiàng)(201302004)
【分類號(hào)】:R123

【相似文獻(xiàn)】

相關(guān)期刊論文 前2條

1 文其乙,劉秀梵,焦新安,Phillip Werner,Boehm Reinhard;多重聚合酶鏈反應(yīng)檢測(cè)污水中產(chǎn)氣莢膜梭菌[J];揚(yáng)州大學(xué)學(xué)報(bào)(自然科學(xué)版);2001年04期

2 孫菊華;麥爾耶姆·薩吾提;;產(chǎn)氣莢膜梭菌:一種水污染的指示菌[J];大家健康(學(xué)術(shù)版);2011年15期

,

本文編號(hào):2145133

資料下載
論文發(fā)表

本文鏈接:http://sikaile.net/yixuelunwen/yufangyixuelunwen/2145133.html


Copyright(c)文論論文網(wǎng)All Rights Reserved | 網(wǎng)站地圖 |

版權(quán)申明:資料由用戶60e9b***提供,本站僅收錄摘要或目錄,作者需要?jiǎng)h除請(qǐng)E-mail郵箱bigeng88@qq.com
日韩黄色一级片免费收看| 欧美一区二区三区不卡高清视| 午夜福利视频日本一区| 国产成人精品一区二区在线看| 91日韩在线视频观看| 亚洲黄色在线观看免费高清| 九九热国产这里只有精品| 丰满人妻少妇精品一区二区三区| 国产亚洲精品岁国产微拍精品| 亚洲av在线视频一区| 国产91色综合久久高清| 国产av天堂一区二区三区粉嫩| 国产大屁股喷水在线观看视频| 国产精品欧美激情在线观看| 欧美多人疯狂性战派对| 美女被啪的视频在线观看| 91一区国产中文字幕| 高清不卡一卡二卡区在线| 欧美一级黄片欧美精品| 国产亚洲系列91精品| 人妻偷人精品一区二区三区不卡| 在线观看国产午夜福利| 国产精品视频一级香蕉| 日本av在线不卡一区| 精品国自产拍天天青青草原| 久久综合九色综合欧美| 久久夜色精品国产高清不卡| 亚洲熟女熟妇乱色一区| 国产亚洲精品岁国产微拍精品| 国产精品免费自拍视频| 欧美日韩在线视频一区| 视频一区二区黄色线观看| 一区二区日韩欧美精品| 国产一区一一一区麻豆| 日韩成人高清免费在线| 老司机这里只有精品视频| 午夜午夜精品一区二区| 亚洲欧美日韩在线看片| 国产在线一区中文字幕| 国产偷拍盗摄一区二区| 久久精品少妇内射毛片|