摘要
建立一种新型的核酸体外等温扩增方法--可视化跨越式滚环等温扩增技术(SRCA)检测沙门氏菌。根据沙门氏菌stn基因序列设计引物,进行特异性验证。测定该技术的灵敏度以及人工污染鸡肉样品的检出限。通过检测多种实际食品样品,评价SRCA方法的敏感性、特异性和符合率。研究结果表明:所有沙门氏菌呈阳性结果,非沙门氏菌呈阴性结果,说明引物特异性良好。SRCA方法检测沙门氏菌DNA的灵敏度为5.6×10^0 fg/μL,检出限为3.8×10^1 CFU/mL;PCR方法的灵敏度为5.6×10^2 fg/μL,检出限为3.8×10^3 CFU/mL。该方法检测30个实际样品的检出率为13.33%,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%,96.30%和96.67%。结论:可视化跨越式滚环等温扩增技术具有操作简便,特异性强,灵敏度高,检出限低,检测成本低,对试验设备要求低等优点,适用于基层单位对食品中沙门氏菌的快速检测。
A new type of vitro nucleic acid isothermal amplification method-saltatory rolling circle amplification(SRCA)was established to detect Salmonella.The primers were designed using the specific gene stn of Salmonella and the specificity was verified.The performance of the assay was evaluated with respect to sensitivity and the detection limit of artificially contaminated chicken was tested.In addition,the sensitivity,specificity and accuracy were evaluated by testing a variety of real food samples.Results:All Salmonella presented positive results,but non-Salmonella gave negative results,indicating good specificity of the primer.The sensitivity of SRCA method was 5.6×10^0 fg/μL for DNA,and the detection limit was 3.8×10^1 CFU/mL.The sensitivity of PCR method was 5.6×10^2 fg/μL,and the detection limit was 3.8×10^3 CFU/mL.In addition,the detection rate of this method for detecting 30 real samples was 13.33%.Compared with the GB 4789.4-2016,the sensitivity,specificity and accuracy of SRCA were 100.00%,96.30%and 96.67%,respectively.Conclusions:The method has the advantages of simple operation,high specificity,high sensitivity,low detection limit,low cost and low equipment requirement,and is suitable for the rapid detection of Salmonella in food.
作者
张蕴哲
苑宁
李靳影
张梦泽
马晓燕
张伟
Zhang Yunzhe;Yuan Ning;Li Jinying;Zhang Mengze;Ma Xiaoyan;Zhang Wei(College of Food Science and Technology,Hebei Agricultural University,Baoding 071001,Hebei;College of Science and Technology,Hebei Agricultural University,Cangzhou 061100,Hebei;College of Life Sciences,Hebei Agricultural University,Baoding 071001,Hebei)
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2020年第9期304-311,共8页
Journal of Chinese Institute Of Food Science and Technology
基金
国家自然科学基金项目(31371772)
河北省自然科学基金项目(C2019204342)
河北省重点研究开发项目(18275501D)。
