Taylor Carl, Lough Fraser, Stanforth Stephen P, Schwalbe Edward C, Fowlis Ian A, Dean John R
Department of Applied Sciences, Northumbria University, Newcastle upon Tyne, NE1 8ST, UK.
Anal Bioanal Chem. 2017 Jul;409(17):4247-4256. doi: 10.1007/s00216-017-0375-x. Epub 2017 May 8.
Listeria monocytogenes is a Gram-positive bacterium and an opportunistic food-borne pathogen which poses significant risk to the immune-compromised and pregnant due to the increased likelihood of acquiring infection and potential transmission of infection to the unborn child. Conventional methods of analysis suffer from either long turn-around times or lack the ability to discriminate between Listeria spp. reliably. This paper investigates an alternative method of detecting Listeria spp. using two novel enzyme substrates that liberate exogenous volatile organic compounds in the presence of α-mannosidase and D-alanyl aminopeptidase. The discriminating capabilities of this approach for identifying L. monocytogenes from other species of Listeria are investigated. The liberated volatile organic compounds (VOCs) are detected using an automated analytical technique based on static headspace-multi-capillary column-gas chromatography-ion mobility spectrometry (SHS-MCC-GC-IMS). The results obtained by SHS-MCC-GC-IMS are compared with those obtained by the more conventional analytical technique of headspace-solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). The results found that it was possible to differentiate between L. monocytogenes and L. ivanovii, based on their VOC response from α-mannosidase activity.
单核细胞增生李斯特菌是一种革兰氏阳性菌,也是一种机会性食源性病原体,由于其感染风险增加以及有可能将感染传播给未出生的胎儿,因此对免疫功能低下者和孕妇构成重大风险。传统的分析方法要么周转时间长,要么缺乏可靠地区分李斯特菌属的能力。本文研究了一种使用两种新型酶底物检测李斯特菌属的替代方法,这两种酶底物在α-甘露糖苷酶和D-丙氨酰氨基肽酶存在的情况下会释放外源性挥发性有机化合物。研究了这种方法从其他李斯特菌种中鉴定单核细胞增生李斯特菌的鉴别能力。使用基于静态顶空-多毛细管柱-气相色谱-离子迁移谱(SHS-MCC-GC-IMS)的自动化分析技术检测释放出的挥发性有机化合物(VOC)。将SHS-MCC-GC-IMS获得的结果与通过更传统的顶空-固相微萃取-气相色谱-质谱(HS-SPME-GC-MS)分析技术获得的结果进行比较。结果发现,根据它们对α-甘露糖苷酶活性的VOC响应,可以区分单核细胞增生李斯特菌和伊氏李斯特菌。