Holmquist Leif, Vesterberg Olof
National Institute for Working Life, Ekelundsvägen 16, Solna, SE-11279 Stockholm, Sweden.
J Biochem Biophys Methods. 2003 Sep 30;57(3):183-90. doi: 10.1016/s0165-022x(03)00107-6.
An immunochromatographic method for qualitative and quantitative determination of aeroallergens direct on sampling (ADOS) filters has been developed. In this method, a porous polytetrafluoroethylene filter carrying adsorbed allergens is fixed by double-coated adhesive tape to a supporting filter paper matrix. Following addition of antibodies specific for the relevant allergens and washing and staining reagents via a reagent applicator an immunochromatogram is developed resulting in a 5-10 mm wide area of the sample filter covered with blue-violet-stained spots appearing on a faintly pink or white background. The method takes 30 to 90 min, depending on the nominal porosity (1.2-5 microm) and the defined reaction area (5-10 mm) of the sample filter. Application experiments with birch and grass pollen, soluble Bet v 1, Phl p 5 and mould allergens as well as cat allergen carried by airborne dust revealed a limit of detection of a few picograms of allergen as stained spots. The specificity of the new method to evaluate the type of allergen is a function of the selected antibodies. The concentrations of the allergen in an air sample are related to the number and intensity of stained spots.
已开发出一种用于在采样(ADOS)滤器上直接定性和定量测定空气变应原的免疫层析方法。在该方法中,将携带吸附变应原的多孔聚四氟乙烯滤器用双面胶带固定在支撑滤纸基质上。通过试剂施加器加入针对相关变应原的抗体以及洗涤和染色试剂后,形成免疫层析图,在淡粉色或白色背景上,样品滤器上会出现5 - 10毫米宽的区域覆盖着蓝紫色染色斑点。该方法需要30至90分钟,具体取决于样品滤器的标称孔隙率(1.2 - 5微米)和定义的反应区域(5 - 10毫米)。对桦树和草花粉、可溶性Bet v 1、Phl p 5和霉菌变应原以及空气中灰尘携带的猫变应原进行的应用实验表明,作为染色斑点的变应原检测限为几皮克。评估变应原类型的新方法的特异性取决于所选抗体。空气样品中变应原的浓度与染色斑点的数量和强度相关。