肠炎沙门氏菌的LAMP检测方法的建立

Detection of Salmonella enteritidis by loop-mediated isothermal amplification

建立肠炎沙门氏菌(Salmonella enteritidis)的环介导等温扩增(Loop-mediated isothermal amplification,LAMP)方法,实现对肠炎沙门氏菌的快速检测。通过针对肠炎沙门氏菌血清型特异性基因lygD设计LAMP内外引物对,优化LAMP扩增反应条件,采用包括肠炎沙门氏菌在内的10种不同菌株进行LAMP引物特异性检测;通过系列梯度稀释肠炎沙门氏菌菌液进行LAMP扩增,计算检出限;并对鲜鸡蛋模拟样本进行LAMP检测。结果表明LAMP法可快速特异地检测出肠炎沙门氏菌;细菌培养液检出限为2.33×101cfu/mL,鲜鸡蛋模拟样品为2.67×101cfu/mL。该方法反应灵敏度高,可用于食品中肠炎沙门氏菌的快速检测。

To detect Salmonella enteritidis rapidly, loop-mediated isothermal amplification （LAMP） method was developed. According to Salmonella enteritidis serotype specific gene lygD, LAMP inner and outer primer sets were designed. Specificity of LAMP primers were validated by assaying 10 different strains. Sensitivity of LAMP method was evaluated by serial gradient dilution of Salmonella enteritidis solution. LAMP used for the detection of egg samples. The detection limits of LAMP assay were 2.33×10^1cfu/ml for pure cultures and 2.67×10^1cfu/ml for analogy egg samples. LAMP method is a simple, rapid and feasible method for the detection of Salmonella enteritidis.