Department of Food Science, Molecular Food Microbiology Laboratory, West Lafayette, IN, USA.
J Appl Microbiol. 2011 Jul;111(1):93-104. doi: 10.1111/j.1365-2672.2011.05040.x. Epub 2011 May 20.
To investigate the suitability of human Hsp60, a receptor for Listeria adhesion protein (LAP), on paramagnetic beads (PMB) to capture Listeria monocytogenes from food in the presence of other Listeria to facilitate rapid and specific detection of this pathogen.
Commercially available streptavidin-coated PMBs were linked with biotinylated Hsp60 (PMB-Hsp60), and the bacterial capture efficiency from pure culture and meat samples was determined. Capture rate was also compared with the monoclonal antibody (MAb)-C11E9-coated beads (PMB-C11E9) and the commercial Dynabeads anti-Listeria. Captured cells were detected and quantified by plating on selective medium, quantitative real-time PCR (qPCR) and a light-scattering sensor. Overall, all ligand-coated beads had similar capture efficiency (varied from 1·8 to 9·2%) for L. monocytogenes under the conditions employed, and the minimum cell number required to achieve such capture was 10³ CFU ml⁻¹. PMB-Hsp60 had significantly greater capture efficiency for pathogenic Listeria (P < 0·0001) than the nonpathogenic Listeria. In contrast, PMB-C11E9 and Dynabeads anti-Listeria had similar capture efficiency for both. The efficacy of all PMBs to capture L. monocytogenes in the presence of Listeria innocua from food matrices was compared. Although Dynabeads anti-Listeria had the overall best capture efficiency, PMB-Hsp60 was able to selectively capture L. monocytogenes even in the presence of 10-100-fold more L. innocua cells from enriched meat samples.
Data show that the human cell receptor, Hsp60, is suitable for the capture of pathogenic Listeria on PMB in the presence of other Listeria in food.
As pathogen interaction with host cells is highly specific, host cell receptors could be used as alternate capture molecules on PMB to aid in specific detection of pathogens.
研究人类热休克蛋白 60（Hsp60）作为李斯特菌黏附蛋白（LAP）受体在顺磁珠（PMB）上的适用性，以捕获食品中的李斯特菌，从而促进对这种病原体的快速和特异性检测。
将商业上可获得的链霉亲和素包被的 PMB 与生物素化的 Hsp60（PMB-Hsp60）连接，并确定从纯培养物和肉样中捕获细菌的效率。还比较了捕获率与单克隆抗体（MAb）-C11E9 包被珠（PMB-C11E9）和商业 Dynabeads 抗李斯特菌的捕获率。通过在选择性培养基上平板计数、实时定量 PCR（qPCR）和光散射传感器来检测和定量捕获的细胞。总体而言，在使用的条件下，所有配体包被珠对李斯特菌的捕获效率（在 1.8 到 9.2%之间变化）相似，达到这种捕获所需的最低细胞数为 10³CFU ml⁻¹。PMB-Hsp60 对致病性李斯特菌的捕获效率明显高于非致病性李斯特菌（P < 0.0001）。相比之下，PMB-C11E9 和 Dynabeads 抗李斯特菌对两者的捕获效率相似。比较了所有 PMB 在食品基质中存在无害李斯特菌时捕获单核细胞增生李斯特菌的效果。虽然 Dynabeads 抗李斯特菌的总体捕获效率最佳，但 PMB-Hsp60 即使在从富含肉样的 10-100 倍更多的无害李斯特菌细胞存在的情况下，也能够选择性地捕获单核细胞增生李斯特菌。
数据表明，人类细胞受体 Hsp60 适合在食品中存在其他李斯特菌的情况下，在 PMB 上捕获致病性李斯特菌。
由于病原体与宿主细胞的相互作用具有高度特异性，因此宿主细胞受体可以用作 PMB 上的替代捕获分子，以辅助对病原体的特异性检测。
