应用环介导等温扩增法快速检测食品中沙门氏菌的研究
发布时间：2018-08-18 10:19
【摘要】：目的建立快速、准确检测食品中沙门氏菌基因的方法。方法针对沙门氏菌高侵袭性位点A(hyper invasive locus A,invA)基因设计4条LAMP引物。提取沙门氏菌基因组DNA,与LAMP反应液及显色液混匀后进行扩增反应,1h内通过颜色变化观察结果。将细菌DNA 10倍系列稀释后分别进行LAMP及PCR反应,评价LAMP法的敏感性。对50份已知食物样品进行检测,评价LAMP法的特异性。结果 LAMP法可在40min内检出沙门氏菌。其敏感性为0.05ng/ml DNA,是PCR方法(0.5ng/ml DNA)的10倍,特异性与PCR方法相当;LAMP法检测食品中沙门氏菌的符合率为98%,PCR法为90%。结论采用建立的LAMP法检测食物样品中的沙门氏菌快速、准确、操作简单,无需要特殊仪器,适合基层相关部门应用。
[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.
【作者单位】： 枣庄矿业集团中心医院;济南市市中区人民医院;
【分类号】：R155.5
[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.
【作者单位】： 枣庄矿业集团中心医院;济南市市中区人民医院;
【分类号】：R155.5
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