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TaqMan探针法荧光定量PCR检测鲜切甜瓜中单核细胞增多性李斯特菌的研究 被引量:3

Study on detection of Listeria monocytogenes in fresh-cut melon with TaqMan-based fluorescence quantitative PCR
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摘要 根据多条单核细胞增多性李斯特菌的hly基因序列,设计了1对简并引物和1条TaqMan探针,建立荧光定量PCR快速检测单核细胞增多性李斯特菌的方法,对该方法的特异性、灵敏度进行了验证,并采用该法对染菌的鲜切甜瓜进行了检测。结果表明,该方法具有较好的特异性,对质粒标准品的灵敏度为1.12×102copies/μL,对染菌的鲜切甜瓜中单核细胞增多性李斯特菌的最低检出限为6.28×102cfu/mL。该方法可为快速检测鲜切果蔬病原微生物污染度及其控制奠定技术基础。 According to the multiple hly gene sequences of Listeria monocytogenes, a pair of degenerate primers and a probe was designed,and the method of real-time fiuorogenic quantitative PCR assay to detection of Listeria monocytogenes was established,the specificity and sensitivity were verified; its preliminary application in artificial inoculation fresh-cut melon was realized.The results showed that the method had good specificity and sensitivity for plasmid standard was 1.12× 10^2copies/μL,the minimum detection limit of artificial inoculation fresh-cut melon was 6.28 × 10^2 cfu/mL. This method can lay a technology foundation to detect and control the pathogenic microorganism of fresh-cut fruits and vegetables.
出处 《食品工业科技》 CAS CSCD 北大核心 2014年第19期297-300,共4页 Science and Technology of Food Industry
基金 国家自然科学基金项目(31172009 31340038) 国家科技支撑计划项目(2012BAD 38B05) 大连市科技计划项目(2012E13SF106) 大连市金州新区科技计划项目(2012-A1-049)
关键词 单核细胞增多性李斯特菌 hly基因 荧光定量PCR TAQMAN探针 鲜切甜瓜 Listeria monocytogenes hly gene real- time fluorogenic quantitative PCR TaqMan probe fresh- cut melon
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