进口鸡翼中单核细胞增生李斯特菌的鉴定及其 挥发性产物分析
目的 对从进口鸡翼中分离的菌株F7-10进行种属鉴定, 并通过顶空气相色谱-质谱联用法(headspace gas chromatography-mass spectrometry, HS-GC-MS)分析其液态培养代谢的挥发性产物。方法 通过VITEK 2 Compact全自动微生物鉴定系统分析菌株的生理生化特征, 结合16S rRNA基因和prfA基因PCR扩增测序鉴定分离菌株, 通过HS-GC-MS分析分离菌株F7-10代谢的挥发性产物, 并与单核细胞增生李斯特菌ATCC 13932、格氏李斯特菌ATCC 700545和英诺克李斯特菌ATCC 33090的代谢挥发性产物进行比较分析。结果 菌株F7-10为革兰氏阳性菌, 生理生化特征与单核细胞增生李斯特菌(Listeria monocytogenes)的相似性为98%, 协同溶血实验在靠近金黄色葡萄球菌的接种端溶血增强; 16S rRNA基因序列与单增李斯特菌标准菌株的相似性为99%, prfA基因特异性扩增出现预期大小的目的片段。菌株F7-10在营养肉汤中代谢的挥发性产物主要有乙醇、3-羟基-2-丁酮、乙酸、丁酸、3-甲基戊酸、十二醇等, 与单增李斯特菌标准菌株的代谢挥发性产物种类相似, 但含量有差别。结论 进口鸡翼中分离的菌株F7-10鉴定为单核细胞增生李斯特菌, 该菌产生的挥发性产物可为单核细胞增生李斯特菌的鉴定提供参考。
Objective To identify and characterize a bacterial strain F7-10 isolated from the imported chicken wings and analyze its volatile compounds (VCs) of liquid-state fermentation by headspace gas chromatography-mass spectrometry (HS-GC-MS). Methods The physiological and biochemical characteristics of the strain were elucidated by using VITEK 2 compact automated microbiology system. The 16S rRNA gene was sequenced and prfA gene from F7-10 was amplified by PCR to detect strain F7-10. Volatile compounds from F7-10 cultured with nutrient broth were detected by HS-GC-MS. The VCs emitted from four strains, including Listeria monocytogenes ATCC 13932, Listeria grayi ATCC 70054, Listeria innocua ATCC33090 and F7-10, were analyzed and compared. Results Strain F7-10 was identified as gram-positive bacteria and exhibited the highest levels of 98% probability to be Listeria monocytogenes based on the conventional physiological test. An enhanced zone of β-haemolysis at the intersection of Staphyloccocus aureus was found. The sequence analysis of 16S rRNA gene of F7-10 isolation showed that it was very close to Listeria monocytogenes and the similarity was 99%. The purposed gene fragment was detected based on prfA gene amplification. The main VCs produced by F7-10 in nutrient broth were detected to be ethanol, acetoin, acetic acid, butanoic acid, 3-methylvaleric acid and dodecyl alcohol. The varieties of VCs behaved similarly between F7-10 and Listeria monocytogenes standard strain, but the content behaved differently. Conclusion Bacterial strain F7-10 isolated from the imported chicken wings is identified as Listeria monocytogenes. The VCs produced by F7-10 can provide basis for the identification of Listeria monocytogenes.
标题：进口鸡翼中单核细胞增生李斯特菌的鉴定及其 挥发性产物分析
英文标题：Identification of Listeria monocytogenes from imported chicken wings and analysis of its volatile compounds
作者：
李正义 山东出入境检验检疫局检验检疫技术中心
贾俊涛 山东出入境检验检疫局检验检疫技术中心
唐静 山东出入境检验检疫局检验检疫技术中心
马云 山东出入境检验检疫局检验检疫技术中心
姜英辉 山东出入境检验检疫局检验检疫技术中心
李兆杰 威海出入境检验检疫局
赵丽青 山东出入境检验检疫局检验检疫技术中心
中文关键词:单核细胞增生李斯特菌,鉴定,挥发性产物,顶空气相色谱-质谱联用法,
英文关键词:Listeria monocytogenes,identification,volatile compounds,headspace gas chromatography-mass spectrometry,
发表日期：2016-11-12
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