叠氮溴化丙锭结合实时荧光定量PCR技术检测发酵乳中植物乳杆菌P-8活菌数

被引：8

作者：

韩之皓

郭帅

黄天

郑岩

王月娇

白梅

王记成

张和平

机构：

[1] 内蒙古农业大学乳品生物技术与工程教育部重点实验室农业部奶制品加工重点实验室

来源：

食品与发酵工业 | 2019年 / 45卷 / 01期

关键词：

植物乳杆菌; 叠氮溴化丙锭(propidium monoazide,PMA); 实时荧光定量PCR;

D O I：

10.13995/j.cnki.11-1802/ts.018232

中图分类号：

TS252.7 [产品标准与检验];

学科分类号：

摘要：

采用叠氮溴化丙锭(propidium monoazide,PMA)结合实时荧光定量PCR对乳制品中活菌DNA进行定量分析,建立一种快速而准确检测发酵乳品中植物乳杆菌P-8(Lactobacillus plantarum P-8)活菌数的新方法。通过对影响PMA作用的浓度、暗孵育和曝光时间等因素进行试验,确定最佳PMA处理方案。结果表明:L. plantarum P-8经80℃处理60 s,即为膜损伤菌;当PMA质量浓度为40μg/m L,暗孵育时间10 min,曝光时间为20 min时,PMA既不影响活菌DNA的PCR扩增,又能渗透进入细胞膜受损的死菌并抑制其PCR扩增;通过制备L.plantarum P-8质粒标准品并建立标准曲线,其表现出良好的线性关系,相关系数(R2)为0. 992 9,最低检测限为103CFU/m L,特异性良好。该方法为完善发酵乳产品益生菌活菌数的检测奠定了基础。

引用

页码：183 / 189

页数：7

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