本文選題：熒光定量PCR + 有毒藍(lán)藻　； 參考：《環(huán)境與健康雜志》2016年02期

【摘要】：目的建立水中有毒藍(lán)藻的熒光定量PCR檢測方法。方法取20 ml純培養(yǎng)藻液和500 ml采集水樣用0.22μm濾膜進(jìn)行過濾,收集藻體提取DNA,應(yīng)用SYBR Green對(duì)微囊藻毒素合成酶基因mcy A進(jìn)行實(shí)時(shí)熒光定量PCR檢測,根據(jù)軟件獲取的以模板標(biāo)準(zhǔn)品與Ct值建立的標(biāo)準(zhǔn)曲線,計(jì)算樣品中的mcy A基因拷貝數(shù)和產(chǎn)微囊藻毒素(MC)的藍(lán)藻的濃度。結(jié)果在13.8~1.38×106拷貝/L的線性范圍內(nèi),所得回歸方程為y=-3.45 lgx+36.2,r=0.989 9。方法檢出限為10.7拷貝/L,加標(biāo)回收率為88.4%~106.0%,RSD為3.6%~4.0%。應(yīng)用該方法和傳統(tǒng)的顯微鏡檢法對(duì)實(shí)驗(yàn)室純培養(yǎng)產(chǎn)毒銅綠微囊藻的檢測結(jié)果呈正相關(guān)(r=0.994 0,P0.05)。結(jié)論該方法靈敏度高、精確度高、檢測范圍寬、重復(fù)性好,適合用于藍(lán)藻水華暴發(fā)的早期預(yù)警。
[Abstract]:Objective to establish a fluorescence quantitative PCR method for the detection of toxic cyanobacteria in water. Methods 20 ml pure culture algae solution and 500ml water sample were filtrated with 0.22 渭 m filter membrane, and the microcystin-synthase gene mcy A was detected by real-time fluorescence quantitative PCR (PCR), and the microcystin synthase gene mcy A was detected by SYBR Green. According to the standard curve obtained by software, the copy number of mcy A gene and the concentration of cyanobacteria producing microcystins were calculated. Results in the linear range of 13.8 ~ 1.38 脳 10 ~ (6) copies / L, the regression equation was obtained as follows: yaw-3.45 lgx ~ (36.2) / L ~ (0.989 9). The detection limit was 10.7 copies 路L ~ (-1) and the recovery rate was 88.4% and 106.0% RSD was 3.6% and 4.0% respectively. There was a positive correlation between this method and the traditional microscope method for the detection of microcystis aeruginosa produced by pure culture in laboratory. Conclusion this method has the advantages of high sensitivity, high accuracy, wide detection range and good repeatability. It is suitable for early warning of Shui Hua outbreaks in cyanobacteria.
【作者單位】： 山東省城市供排水水質(zhì)監(jiān)測中心;山東大學(xué)環(huán)境科學(xué)與工程學(xué)院;
【基金】：住房與城鄉(xiāng)建設(shè)部國家水體污染控制與治理科技重大專項(xiàng)(2012ZX07404-003) 國家科技惠民工程(2013GS370202) 山東省教育廳泰山學(xué)者建設(shè)工程專項(xiàng)(ts200640025)
【分類號(hào)】：R123.1;R440

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