Peters Marcus, Bonowitz Petra, Bufe Albrecht
Department of Experimental Pneumology, Ruhr University Bochum, Bochum, Germany.
Methods Mol Biol. 2017;1600:143-150. doi: 10.1007/978-1-4939-6958-6_14.
The availability of convenient assays for the detection and quantification of pathogen-associated molecular patterns (PAMPs) is limited. In the case of lipopolysaccharide (LPS) the so-called LAL (limulus amebocyte lysate) test is available, an assay that is performed with the lysate of the blood of the horse shoe crab. Although a sensitive and convenient assay, it lacks specificity, since it is affected by other endotoxins like, for instance, fungal cell walls as well. Here, we describe a bioassay that can be used to detect and quantitate PAMPs in environmental samples. More specific we demonstrate the usage of TLR2 and TLR4/CD14/MD2 transfected Hek293 cells to quantitatively determine bacterial lipoproteins and LPS, respectively. We show the usefulness of these assays to measure LPS in tobacco before and after combustion.
用于检测和定量病原体相关分子模式(PAMP)的便捷检测方法有限。就脂多糖(LPS)而言,有所谓的鲎试剂(LAL,鲎血细胞溶解物)检测法,该检测是用鲎的血液裂解物进行的。尽管这是一种灵敏且便捷的检测方法,但它缺乏特异性,因为它也会受到其他内毒素的影响,例如真菌细胞壁。在此,我们描述了一种可用于检测和定量环境样品中PAMP的生物检测法。更具体地说,我们展示了用转染了TLR2和TLR4/CD14/MD2的Hek293细胞分别定量测定细菌脂蛋白和LPS的方法。我们证明了这些检测方法在测量烟草燃烧前后LPS方面的实用性。