Fluorescence immunoassay using an optical fiber for determination of Dermatophagoides farinae (Der f1)
Fluorescence immunoassay using an optical fiber for determination of Dermatophagoides farinae (Der f1)
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DOI：
10.1007/s10661-011-1872-6
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发表时间：
2011-01
影响因子：
3
通讯作者：
K. Miyajima;Gen Itabashi;Tomoyuki Koshida;K. Tamari;D. Takahashi;T. Arakawa;H. Kudo;H. Saito;K. Yano;K. Shiba;K. Mitsubayashi
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文献类型：
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作者：
K. Miyajima;Gen Itabashi;Tomoyuki Koshida;K. Tamari;D. Takahashi;T. Arakawa;H. Kudo;H. Saito;K. Yano;K. Shiba;K. Mitsubayashi
免疫测定方法通常用于测量过敏性物质。然而，它们需要特殊的设施、熟练的操作和耗时的程序。本工作建立了一种光纤免疫分析系统，该系统可以通过过敏原与荧光标记抗体形成的免疫复合物的荧光强度来检测过敏原。免疫复合物吸附在光纤探针表面，激发光注入光纤探针，在光纤探针附近产生倏逝场。荧光团由渐逝光激发，并且荧光由光电二极管检测。本系统检测的目标过敏原为粉尘螨Dermatophagoides farinae的Der f1，粉尘螨是屋尘螨之一，也是吸入性过敏原的主要来源。用于检测抗体的标记荧光团为花青5。该系统能够检测和定量测定ofDer f1。测定范围为0.24 ~ 250 ng/ml，结果与ELISA竞争。测量时间为16分钟/样品。将该免疫分析系统应用于实际粉尘样品中Der f1的测定。Der f1的计算值与ELISA法具有良好的相关性。
Immunoassay methods are generally used for measuring of allergenic substances. However, they need special facilities, skilled handling, and time-consuming procedure. In this work, a fiber-optic immunoassay system which could measure allergen by fluorescent intensities of immune complexes formed by allergens and fluorescently labeled antibodies was established. Immune complexes absorbed on the optical fiber probe surface, and excitation light was injected into the probe, then evanescent field is created in the proximity of the probe. The fluorophores were excited by the evanescent light, and fluorescence was detected by a photo diode. The target allergen detected by our system wasDer f1 derived fromDermatophagoides farinaethat is one of the house dust mite and major source of inhaled allergens. The fluorophore used labeling on detecting antibody was cyanine 5. The system enabled to detect and quantitatively determine ofDer f1. The measurement range was from 0.24 to 250 ng/ml, and the result competes with ELISA. The measurement time was 16 min/sample. The immunoassay system was applied to measurement ofDer f1 from actual dust samples. Calculated values ofDer f1 showed good correlations between the fiber-optic fluoroimmunoassay and ELISA.