免疫磁珠分离-实时荧光PCR快速检测虾中沙门氏菌

被引:16
作者
李亚茹 [1 ]
周冬根 [2 ]
夏杏洲 [3 ]
曹怡芳 [1 ]
杨慧宁 [1 ]
胡双芳 [1 ]
肖性龙 [1 ]
机构
[1] 华南理工大学食品科学与工程学院
[2] 宁波检验检疫科学技术研究院
[3] 广东海洋大学食品科技学院
基金
浙江省自然科学基金; 广东省自然科学基金; 广东省科技计划;
关键词
免疫磁珠分离; 实时荧光PCR; 沙门氏菌; 检测;
D O I
10.13982/j.mfst.1673-9078.2017.11.034
中图分类号
TS254.7 [水产制品的标准与检验];
学科分类号
摘要
建立了免疫磁珠分离(Immunomagnetic separation,IMS)联合实时荧光PCR(real time PCR)快速、灵敏地检测虾中沙门氏菌的方法。用纳米磁珠与抗沙门氏菌多克隆抗体制备免疫磁珠,优化反应条件,建立IMS方法。同时针对沙门氏菌ttr基因合成探针和引物,构建real time PCR体系,选4株代表性的沙门氏菌检测其特异性和灵敏度。结果显示,免疫磁珠的最适添加量为100μL,免疫磁珠与样品菌液的最佳反应时间为30 min。建立的免疫磁珠分离-实时荧光PCR(IMS-real time PCR)方法特异性高,对于虾中沙门氏菌的检测限为鼠伤寒沙门氏菌5×10~1 CFU/25 g,猪霍乱沙门氏菌5×10~2 CFU/25 g,肠炎沙门氏菌和甲型副伤寒沙门氏菌1×10~2CFU/25 g,检测全程可在6 h内完成。对40份实际样品,IMS-real time PCR方法与国标法检测结果完全一致。建立的IMS-Real time PCR方法用时短、灵敏度高,为水产品中沙门氏菌的快速检测提供了技术支撑,对沙门氏菌引起的食源性疾病的预防和控制具有现实意义。
引用
收藏
页码:235 / 242
页数:8
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