摘要
建立了可视化单引物等温扩增技术(Visual single primer isothermal amplification,Visual SPIA)检测发酵乳制品中沙门氏菌(Salmonella)的方法。针对沙门氏菌侵袭蛋白(Invasion protein A,invA)基因设计引物,验证该方法特异性,同时对人工污染发酵乳制品的检出限进行测定。分别采用国家标准方法(GB 4789.4—2016)和可视化SPIA方法对235份发酵乳制品样品进行检测,确定可视化SPIA方法的敏感性、特异性及符合率。结果显示,12株沙门氏菌呈阳性结果,29株非沙门氏菌均呈阴性结果。荧光可视法的检出限均为3.2×10^(1) CFU/mL,沉淀可视法的检出限为3.2×10^(2) CFU/mL。可视化SPIA方法的敏感性为100.00%,特异性为98.15%,符合率为99.15%。该方法特异性强,可通过肉眼直接观察并判定结果,实现了对沙门氏菌的可视化检测,对沙门氏菌引起的食物中毒的预防和控制具有重要意义。
In this study,a visual single primer isothermal amplification(visual single primer isothermal amplification,visual SPIA)method was established to detect Salmonella in fermented dairy products.Primers were designed targeting Salmonella invasion protein(Invasion protein A,invA)gene to verify the specificity of the method.The detection limit of artificially contaminated fermented dairy products was determined.To ascertain the sensitivity,specificity and accuracy of the visual SPIA,the national standard method(GB 4789.4—2016)and the visual SPIA method were used to detect 235 samples of fermented dairy products.The results revealed that 12 strains of Salmonella presented positive results and 29 strains of non-Salmonella showed negative results.The detection limit by fluorescence was 3.2×10^(1) CFU/mL and the detection limit by precipitation was 3.2×10^(2) CFU/mL.The sensitivity,specificity and accuracy of the visual SPIA method were 100.00%,98.15%and 99.15%,respectively.The method with wonderful specificity can directly observe and judge the results by the naked eye,realizing the visual detection of Salmonella.In all,the visual SPIA method was of great significance to the prevention and control of food poisoning caused by Salmonella.
作者
王妙姝
范春刚
郑玮丽
王震
马晓燕
WANG Miaoshu;FAN Chun'gang;ZHENG Weili;WANG Zhen;MA Xiaoyan(New Hope Tensun(Hebei)Dairy Co.,Ltd.,Baoding,Hebei 072159,China;College of Food Science and Technology,Hebei Agricultural University,Baoding,Hebei 071001,China;Hebei Province Probiotic Functional Dairg Product Technology Innovation Center,Baoding,Hebei 072159,China)
出处
《农产品加工》
2021年第17期53-56,共4页
Farm Products Processing
基金
保定市科技计划项目(18FH05)。
关键词
可视化
单引物等温扩增
沙门氏菌
检测
发酵乳制品
visual
single primer isothermal amplification
Salmonella
detection
fermented dairy products
