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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。
Objective To establish a rapid and accurate method for detecting Salmonella in food. Methods Four LAMP primers were designed according to the invA gene of Salmonella. Salmonella genomic DNA was extracted and mixed with LAMP reaction solution and chromogenic solution for amplification reaction. The color change was observed within 1 hour. After 10-fold serial dilution of bacterial DNA, LAMP and PCR reactions were respectively performed to evaluate the sensitivity of LAMP method. 50 samples of known food were tested to evaluate the specificity of the LAMP method. Results LAMP method can detect Salmonella within 40 min. Its sensitivity is 0.05ng / ml DNA, which is 10 times higher than that of PCR method (0.5ng / ml DNA). The specificity is similar to that of PCR method. The coincidence rate of LAMP method for Salmonella in food is 98% and PCR method is 90%. Conclusion The detection of Salmonella in food samples by the established LAMP method is rapid, accurate and easy to operate. It does not require special instruments and is suitable for the application of grassroots units.