IMSA技术快速检测沙门氏菌方法的建立及应用

Establishment and application of IMSA technology in the rapid detection on Salmonella

[目的]建立和评价一种快速、准确地检测食品中沙门氏菌的检测方法,对食品安全控制具有重要意义。[方法]应用等温多自配引发扩增技术(Isothermal Multiple Self-matching-initiated Amplification,IMSA),设计筛选出沙门氏菌的特异性基因invA基因的IMSA引物,并建立检测沙门氏菌的IMSA法。对建立的方法进行特异性和灵敏度试验;确定人工污染样本能被该方法检出的最短增菌时间和最低检测灵敏度;利用IMSA法和培养法对13份食品样本进行检测,评价两者的一致性。[结果]该方法仅对沙门氏菌特异性扩增,灵敏度达3.02×102 CFU/mL;人工污染样本最短培养6 h可被IMSA法检出,检出限为3.17CFU/25 g;13份食品样本的IMSA法与培养法结果一致性为100%。[结论]IMSA技术检测沙门氏菌的灵敏度高、特异性强、结果准确,可在7h内完成食品中沙门氏菌的检测,适合用于食品中沙门氏菌的快速检测。

[Objective]This study establishes and evaluates a rapid and accurate method for detecting Salmonella in food,which is of great significance for control and detection of food safety.[Method]Isothermal Multiple Self-matching-initiated Amplification(IMSA)was used to design primers to detect the specific invA gene of Salmonella and to establish an IMSA method for detecting Salmonella.Perform specificity and sensitivity tests on the established method,and determine the minimum growth time and minimum detection sensitivity of the method for detecting artificially contaminated samples;Thirteen food samples were tested using IMSA and culture methods to evaluate the consistency of the two methods.[Result]This method only specifically amplified Salmonella,and its sensitivity reached 3.02×102 CFU/mL.The shortest incubation time for the IMSA method to detect artificially contaminated samples was 6 hours,and the detection limit was 3.17CFU/25 g.The consistency of IMSA and culture results of thirteen food samples was 100%.[Conclusion]The detection of Salmonella by IMSA technology has the characteristics of high sensitivity,strong specificity and accurate results.It can complete the detection of Salmonella in food within 7 hours,and can be used for rapid detection of Salmonella in food.