本文選題：恒溫實(shí)時(shí)熒光法 + 阪崎腸桿菌�。� 參考：《現(xiàn)代食品科技》2016年09期

【摘要】：為實(shí)現(xiàn)奶粉中快速檢測(cè)阪崎腸桿菌,本文建立了檢測(cè)阪崎腸桿菌的恒溫實(shí)時(shí)熒光法。針對(duì)阪崎腸桿菌16S r RNA設(shè)計(jì)三組LAMP引物,采用Deaou-308C恒溫實(shí)時(shí)熒光檢測(cè)平臺(tái),選取常見(jiàn)病原菌標(biāo)準(zhǔn)株進(jìn)行引物特異性檢測(cè);選取阪崎腸桿菌標(biāo)準(zhǔn)菌株進(jìn)行基因組DNA靈敏度和最低檢測(cè)限測(cè)定,同時(shí)利用人工污染方式檢測(cè)此方法在脫脂和全脂奶粉中的靈敏度和最低檢測(cè)限,利用Real Amp法和國(guó)標(biāo)法對(duì)20份市售奶粉進(jìn)行對(duì)比實(shí)驗(yàn)。結(jié)果顯示,引物組16S-11擴(kuò)增效率最優(yōu),與常見(jiàn)病原菌無(wú)交叉反應(yīng),對(duì)阪崎腸桿菌基因組DNA、阪崎腸桿菌污染的脫脂和全脂奶粉的靈敏度分別達(dá)到102 CFU/m L、102 CFU/m L和103 CFU/m L;對(duì)阪崎腸桿菌基因組DNA和阪崎腸桿菌污染的脫脂和全脂奶粉的最低檢測(cè)限分別達(dá)到103 CFU/m L、103 CFU/m L和104 CFU/m L;在20份市售奶粉樣品中Real Amp檢測(cè)結(jié)果與傳統(tǒng)國(guó)標(biāo)培養(yǎng)結(jié)果一致,表明本文建立的阪崎腸桿菌Real Amp檢測(cè)方法適用于阪崎腸桿菌的快速檢測(cè)。
[Abstract]:In order to rapidly detect Enterobacter sakazakii in milk powder, a real-time isothermal fluorescence method was developed for the detection of Enterobacter sakazakii. Three sets of LAMP primers were designed for Enterobacter sakazakii 16s r RNA. The standard strains of common pathogenic bacteria were selected for specific detection using Deaou-308C real-time fluorescence detection platform. Enterobacter sakazakii standard strains were selected to detect the sensitivity and minimum detection limit of genomic DNA, and the sensitivity and minimum detection limit of this method in defatted and full-fat milk powder were detected by artificial contamination. Real Amp method and national standard method were used to compare 20 samples of milk powder sold in the market. The results showed that the primer group had the best amplification efficiency and had no cross reaction with common pathogens. The sensitivity to Enterobacter sakazakii genome DNA, Enterobacter sakazakii contaminated defatted milk powder and whole milk powder were 102 CFU/m L ~ (10 ~ 2) CFU/m L and 10 ~ 3 CFU/m / L, respectively, while the defatted and whole milk powder contaminated by Enterobacter sakazakii genome DNA and Enterobacter sakazakii were the most sensitive. The low detection limit was 103 CFU/m / L 103 CFU/m / L and 104 CFU/m / L, respectively, and the results of Real Amp detection in 20 samples of marketable milk powder were consistent with those of the traditional national standard. This method is suitable for rapid detection of Enterobacter sakazakii.
【作者單位】： 華南理工大學(xué)輕工與食品學(xué)院;廣州迪澳生物科技有限公司;
【基金】：中央高校基本科研業(yè)務(wù)費(fèi)專項(xiàng)資金資助(2015ZM063) 中國(guó)博士后科學(xué)基金(2015M580723)
【分類號(hào)】：R155.5

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