【摘要】：目的建立快速、準(zhǔn)確檢測(cè)食品中沙門氏菌基因的方法。方法針對(duì)沙門氏菌高侵襲性位點(diǎn)A(hyper invasive locus A,invA)基因設(shè)計(jì)4條LAMP引物。提取沙門氏菌基因組DNA,與LAMP反應(yīng)液及顯色液混勻后進(jìn)行擴(kuò)增反應(yīng),1h內(nèi)通過(guò)顏色變化觀察結(jié)果。將細(xì)菌DNA 10倍系列稀釋后分別進(jìn)行LAMP及PCR反應(yīng),評(píng)價(jià)LAMP法的敏感性。對(duì)50份已知食物樣品進(jìn)行檢測(cè),評(píng)價(jià)LAMP法的特異性。結(jié)果 LAMP法可在40min內(nèi)檢出沙門氏菌。其敏感性為0.05ng/ml DNA,是PCR方法(0.5ng/ml DNA)的10倍,特異性與PCR方法相當(dāng);LAMP法檢測(cè)食品中沙門氏菌的符合率為98%,PCR法為90%。結(jié)論采用建立的LAMP法檢測(cè)食物樣品中的沙門氏菌快速、準(zhǔn)確、操作簡(jiǎn)單,無(wú)需要特殊儀器,適合基層相關(guān)部門應(yīng)用。
[Abstract]:Objective to establish a rapid and accurate method for the detection of Salmonella gene in food. Methods four LAMP primers were designed for the highly invasive A (hyper invasive locus inva gene of Salmonella. The genomic DNA of Salmonella was extracted and mixed with LAMP reaction solution and chromogenic solution. The results of color change were observed within 1 h after amplification reaction. The sensitivity of LAMP method was evaluated by LAMP and PCR reaction after dilution of bacterial DNA 10-fold series. The specificity of LAMP method was evaluated by detecting 50 known food samples. Results Salmonella could be detected by LAMP in 40min. The sensitivity of the method was 10 times that of the PCR method (0.5ng/ml DNA), and its specificity was comparable to that of the PCR method. The coincidence rate of lamp assay for the detection of salmonella in food was 98% and 90% respectively. Conclusion the established LAMP method for the detection of salmonella in food samples is rapid, accurate, easy to operate and does not need special instruments. It is suitable for basic level departments.
【作者單位】： 棗莊礦業(yè)集團(tuán)中心醫(yī)院;濟(jì)南市市中區(qū)人民醫(yī)院;
【分類號(hào)】：R155.5

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