EMA结合实时荧光PCR方法检测单核细胞增生李斯特氏菌

被引：4

作者：

吴海江 ^{[1
]}

孙玉萍 ^{[2
]}

范田丽 ^{[1
]}

赵建勇 ^{[1
]}

张煌涛 ^{[1
]}

张晓波 ^{[2
]}

杨珊珊 ^{[2
]}

机构：

[1] 新疆维吾尔自治区产品质量监督检验研究院/国家农副产品质量监督检验中心

[2] 新疆医科大学基础医学院

来源：

食品与生物技术学报 | 2020年 / 39卷 / 11期

关键词：

单核细胞增生李斯特氏菌; hly基因; 叠氮溴化乙锭; 实时荧光PCR;

D O I：

暂无

中图分类号：

TS207.4 [食品的微生物检验];

学科分类号：

083201 ;

摘要：

建立叠氮溴化乙锭(EMA)结合实时荧光PCR(qPCR)方法检测单核细胞增生李斯特氏菌(Listeria monocytogenes)活菌。以李斯特溶血素O(LLO)基因hly设计引物、TaqMan探针,用李斯特属典型菌、沙门氏菌等56株致病菌株验证特异性,不同质量浓度EMA处理进行qPCR检测。尝试脱氧胆酸钠溶液(SD)强化抑制效果,73份不同的人工污染食品、环境样本(卤鸡肉、牛奶、肉馅、垃圾渗滤液)测试实用性。结果表明,引物探针准确检测L. monocytogenes,对其他菌株无特异性扩增。EMA最适质量浓度2.5μg/mL,经过15 min光激活与死菌DNA共价结合明显抑制了扩增,方法检出限为150 CFU/mL。SD处理L. monocytogenes活菌Ct值增加。与传统培养法比较,EMA-qPCR方法检测100%准确,操作简单、省时高效,在食品、环境方面应用前景广阔。

引用

页码：65 / 70

页数：6

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