不同食品基质中单核细胞增生李斯特氏菌核酸提取方法的比较
Comparative Study of Nucleic Acid Extraction Method of L.monocytogenes in the Different Kinds of Food Matrix
DOI：10.3969/j.issn.1673-1689.2013.11.014
中文关键词: 单核细胞增生李斯特氏菌 食品基质 核酸提取 实时荧光PCR
英文关键词: L.monocytogenes food matrix the extraction of nucleic acid real-time fluorescent PCR
基金项目:国家质检总局科研基金项目(2012IK168)
作者
单位
周阳
安徽农业大学茶与食品科技学院,安徽合肥230000
安徽省食品安全分析与检测省级实验室,安徽合肥230000
祝长青
江苏出入境检验检疫局,江苏南京,210001
郭云昌
国家食品安全风险评估中心,北京,100022
郭桂萍
蒋原
刘秀梅
付瑞燕
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中文摘要:
比较4类食品基质中单核细胞增生李斯特氏菌核酸提取方法的差异。以不同食品基质中的单核细胞增生李斯特氏菌作为研究对象,选取应用广泛并且原理不同的4种细菌核酸提取方法进行比较,同时结合实时荧光PCR法对各种方法进行实际评估。天根离心柱法核酸提取结果稳定,受不同食品基质的影响较小;Promega沉降法与磁珠法受不同食品基质影响较大,核酸提取结果差异较大;Chelex-100法快速方便,但仅适于纯菌液或菌落的核酸提取。
英文摘要:
Compare the methods of extracting nucleic acid of Listeria monocytogenes in four food matrices. Taking L.monocytogenes as our research object,we picked and tested four widely applicable nucleic acid extraction methods and evaluated their performance by real-time PCR.Results:Tiangen centrifugal column method was very stable,with minimum variation caused by different food matrices. Promega precipitation and magnetic bead methods were more affected by food matrices than others,with a larger variation in final products;Chelex-100 method was quick and convenient,but was only suitable for pure liquid culture or colonies.
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