EMA-PCR方法檢測飲用水中3種食源性致病菌活菌研究
發(fā)布時間：2019-05-20 12:18
【摘要】：目的將EMA(Ethidium Monoazide Bromide)選擇滲透性與PCR技術(shù)相結(jié)合,建立有效快速檢測飲用水中3種食源性致病菌活菌EMA-PCR方法。方法以鼠傷寒沙門菌inv A、腸出血性大腸桿菌O157:H7 rfb E、銅綠假單胞菌opr I基因為PCR檢測靶基因,純培養(yǎng)物提取模板進(jìn)行PCR,進(jìn)行EMA使用濃度及靈敏度試驗。結(jié)果 EMA濃度不大于10μg/ml對活菌DNA擴(kuò)增沒有明顯抑制,終濃度為1μg/ml EMA能有效抑制死菌擴(kuò)增;對鼠傷寒沙門菌、腸出血性大腸桿菌O157:H7、銅綠假單胞菌的EMA-PCR靈敏度分別為1.2×104CFU/ml、2×105CFU/ml、3×104CFU/ml;用10μg/ml EMA處理飲用水水樣,可同時檢測出飲用水中含有的3種食源性致病菌活菌。結(jié)論 EMA-PCR方法可一次檢測飲用水中3種食源性致病菌活菌,能避免PCR檢測可能造成假陽性結(jié)果。
[Abstract]:Objective to establish an effective and rapid EMA-PCR method for the detection of three kinds of food-borne pathogenic bacteria in drinking water by combining EMA (Ethidium Monoazide Bromide) selective permeability with PCR technique. Methods Salmonella typhimurium inv A, enterohemorrhagic Escherichia coli O157:H7 rfb E and Pseudomonas aeruginosa opr I gene were used as target genes for PCR detection. PCR, was extracted from pure culture template for EMA concentration and sensitivity test. Results when the concentration of EMA was not more than 10 渭 g / ml, the DNA amplification of living bacteria was not significantly inhibited, but the final concentration of 1 渭 g / ml EMA could effectively inhibit the amplification of dead bacteria. The EMA-PCR sensitivity of Salmonella typhimurium, enterohemorrhagic Escherichia coli O157 and Pseudomonas aeruginosa were 1.2 脳 10 ~ 4 CFU / ml, 2 脳 10 ~ 5 CFU / ml, 3脳104CFU / ml;, respectively. Three kinds of living bacteria containing food-borne pathogenic bacteria in drinking water can be detected at the same time when 10 渭 g / ml EMA is used to treat drinking water samples. Conclusion EMA-PCR method can detect three kinds of living bacteria of food-borne pathogenic bacteria in drinking water at one time, which can avoid the false positive results of PCR detection.
【作者單位】： 南京醫(yī)科大學(xué);江蘇省昆山市疾病預(yù)防控制中心;江蘇省蘇州市疾病預(yù)防控制中心;
【基金】：蘇州市飲用水安全與水性疾病監(jiān)測公共服務(wù)平臺2012年開放課題(SZPT2012002)
【分類號】：R155.5
[Abstract]:Objective to establish an effective and rapid EMA-PCR method for the detection of three kinds of food-borne pathogenic bacteria in drinking water by combining EMA (Ethidium Monoazide Bromide) selective permeability with PCR technique. Methods Salmonella typhimurium inv A, enterohemorrhagic Escherichia coli O157:H7 rfb E and Pseudomonas aeruginosa opr I gene were used as target genes for PCR detection. PCR, was extracted from pure culture template for EMA concentration and sensitivity test. Results when the concentration of EMA was not more than 10 渭 g / ml, the DNA amplification of living bacteria was not significantly inhibited, but the final concentration of 1 渭 g / ml EMA could effectively inhibit the amplification of dead bacteria. The EMA-PCR sensitivity of Salmonella typhimurium, enterohemorrhagic Escherichia coli O157 and Pseudomonas aeruginosa were 1.2 脳 10 ~ 4 CFU / ml, 2 脳 10 ~ 5 CFU / ml, 3脳104CFU / ml;, respectively. Three kinds of living bacteria containing food-borne pathogenic bacteria in drinking water can be detected at the same time when 10 渭 g / ml EMA is used to treat drinking water samples. Conclusion EMA-PCR method can detect three kinds of living bacteria of food-borne pathogenic bacteria in drinking water at one time, which can avoid the false positive results of PCR detection.
【作者單位】： 南京醫(yī)科大學(xué);江蘇省昆山市疾病預(yù)防控制中心;江蘇省蘇州市疾病預(yù)防控制中心;
【基金】：蘇州市飲用水安全與水性疾病監(jiān)測公共服務(wù)平臺2012年開放課題(SZPT2012002)
【分類號】：R155.5
【參考文獻(xiàn)】
相關(guān)期刊論文 前7條
1 張淑紅;吳清平;徐曉可;張菊梅;潘寶怡;彭飛艇;;桶裝水中銅綠假單胞菌檢測方法的比較[J];現(xiàn)代食品科技;2011年11期
2 趙素君;謝晶;曹冶;李江凌;王秋實;葉勇剛;羅丹丹;葉健強;廖黨金;;腸出血性大腸桿菌O157∶H7 EMA-PCR檢測技術(shù)的建立[J];中國草食動物科學(xué);2012年04期
3 馬群飛,林堅,陳美蘭,楊明金,阮國洪;飲用天然礦泉水水源銅綠假單胞菌污染調(diào)查[J];環(huán)境與健康雜志;2001年03期
4 趙瑜;李金磊;董鵬;狄元冉;高延玲;;EMA與PCR結(jié)合檢測沙門氏菌方法的研究[J];畜牧與飼料科學(xué);2013年11期
5 李正義;賈俊濤;祝素珍;姜英輝;房保海;雷質(zhì)文;;細(xì)菌活的非可培養(yǎng)狀態(tài)的分子生物學(xué)檢測技術(shù)研究進(jìn)展[J];食品科學(xué);2011年15期
6 潘樂;孟慶峰;錢愛東;王偉利;;VBNC狀態(tài)細(xì)菌檢測方法的研究進(jìn)展[J];中國農(nóng)學(xué)通報;2012年05期
7 劉U，
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