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。
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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
Abstract: 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 fluorogenic 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 × 102 copies /μL, the minimum detection limit of artificial inoculation fresh-cut melon was 6.28 × 102 cfu /mL. This method can lay a technology foundation to detect and control the pathogenic microorganism of fresh-cut fruits and vegetables. -
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