实时荧光环介导等温扩增技术检测糕点中沙门氏菌的研究

Study on real-time fluorescence loop-mediated isothermal amplification (RF-LAMP) technology for detection of Salmonella in pastry

研究建立一种实时荧光环介导等温扩增技术(Real-time?uorescence loop-mediated isothermal amplification,RF﹣LAMP)检测沙门氏菌的方法。利用invA基因设计引物,进行特异性验证,并对灵敏度和人工污染蛋糕的检出限进行测定。结果显示,沙门氏菌呈阳性结果,非沙门氏菌呈阴性结果,表明特异性良好。该方法检测纯培养物的灵敏度为6.3 cfu/mL,人工污染蛋糕样品的检出限为63 cfu/g。另外,利用该方法对65份糕点样品进行检测,并与GB 4789.4—2016方法进行比较,其敏感性、特异性和符合率分别为100%、98.44%和98.46%。该方法特异性好,灵敏度高,操作简单,可实时监测扩增反应,对沙门氏菌引起的食源性疾病的预防和控制具有重要意义。

This study aimed to establish a real-time fluorescence loop-mediated isothermal amplification(RF-LAMP)assay to detect Salmonella.The primers of Salmonella were designed based on the invA gene.The performance of the assay was evaluated with respect to specificity and sensitivity,and the detection limit of artificially contaminated cake was tested.All Salmonella strains were identified as positive,however,non-Salmonella strains were negative,which showed great specificity.The sensitivity in pure culture by RF-LAMP was 6.3 cfu/mL and detection limit was also evaluated with artificially contaminated cake,and it was 63 cfu/g.In addition,a total of 65 pastry samples were investigated for Salmonella contamination using RF-LAMP assay,and the results were 100.00%sensitivity,98.44%specificity and 98.46%accuracy compared with GB 4789.4—2016 method.The method has high specificity and sensitivity and allows real-time monitoring,which has significant importance for the prevention and control of food-borne diseases caused by Salmonella.