食品中沙门氏菌FTA膜结合跨越式滚环等温扩增检测方法的建立

为实现食品中沙门氏菌的简便和快速现场检测,本研究采用FTA膜(Flinders technology associates,FTA)结合跨越式滚环等温扩增(Saltatory rolling circle amplification,SRCA)方法(FTA-SRCA)建立一种新型的沙门氏菌检测方法.利用FTA膜快速提取...

Development of Detection Method of Salmonella in Food by Saltatory Rolling Circle Amplification Combined with FTA Card

In order to achieve the simple and rapid on-site detection of Salmonella in food, saltatory rolling circle amplification (SRCA) method combined with FTA card (FTA-SRCA) was used to establish a new detection method for Salmonella. The template DNA was extracted by FTA card quickly. According to the specific invA gene of Salmonella, primers were designed and screened. Based on the above, the FTA-SRCA reaction system was established. The reaction was amplificated in the well plate which can achieve intensification. After the reaction, fluorescent dyes were added to observe the results. Its specificity, sensitivity and detection limit of artificial contaminated samples were determined. Furthermore, 60 actual samples were tested to evaluate relative sensitivity, specificity and accuracy. The results indicated that the 17 strains of Salmonella tested were all positive, and the 29 strains of non-Salmonella were negative. Therefore, the method had good specificity. The sensitivity of the FTA-SRCA was 6.81×100 CFU/mL, which was 100 times that of the PCR method and 10 times that of the SRCA method For the detection of artificially contaminated milk samples, the detection limit of the FTA-SRCA was 3.22×100 CFU/mL, which was 1000 times that of the PCR method and 10 times that of the SRCA method. The sensitivity, specificity and accuracy of FTA-SRCA were 100.00%, 94.64% and 95.00%, respectively. The FTA-SRCA method established in this study had the advantages of simple and rapid operation, low cost, strong specificity, high sensitivity, and low detection limit, which can be used for the intensive and rapid on-site detection of Salmonella in food.