Comparative studies of magnetic particle-based solid phase fluorogenic and electrochemiluminescent immunoassay
Comparative studies of magnetic particle-based solid phase fluorogenic and electrochemiluminescent immunoassay
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DOI：
10.1016/s0022-1759(98)00047-7
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发表时间：
1998-09-01
影响因子：
2.2
通讯作者：
Yu, H
中科院分区：
文献类型：
--
作者：
Yu, H
两种固相免疫测定法，电化学发光免疫测定法（ECLIA）和磁颗粒荧光免疫测定法（MPFIA）对细菌检测进行了评估和比较。简而言之，ECLIA是基于钌(II)-三联吡啶Ru[(bpy)(3)]（2+）标记抗体与过量的三丙胺之间的氧化还原反应，产生光子。整个反应是在球形磁珠和阳极电极之间的近表面区域进行的。在ECLIA中，可检测到的细菌孢子在5 × 10(3)至5 × 10(5)个菌落形成单位（cfu）的枯草芽孢杆菌黑变芽孢，10(2)至10(4)个菌落形成单位（cfu）的炭疽芽孢和10(2)至10(6)个大肠杆菌O157:H7细胞之间呈线性范围。独特的MPFIA技术采用抗体包被磁珠作为固相悬浮液，在96孔微孔板中进行细菌捕获和浓度。首先捕获抗体，细菌与碱性磷酸酶（AP）标记的抗体形成夹心，作为报告蛋白，然后与碱性磷酸酶底物atophos （TM）反应，产生荧光用于检测。免疫磁分离允许直接分离和浓缩细菌细胞从原油样品，如血液和环境水。与ECLIA相比，MPFIA检测细菌的灵敏度较低，但它为从粗生物样品中直接、高通量筛选细菌提供了一种手段。ECLIA和MPFIA都是快速（不到一小时）且易于使用的。(C) 1998 Elsevier Science B.V.版权所有
Two solid phase immunoassays, an electrochemiluminescent immunoassay (ECLIA) and a magnetic particle fluorogenic immunoassay (MPFIA) were evaluated and compared for bacterial detection. Briefly, the ECLIA is based on a redox reaction between ruthenium (II)-trisbipyridyl Ru[(bpy)(3)](2+) labeled antibody and the excess of tripropylamine, which generates photons. The entire reaction is carried on the near surface area between the spherical magnetic beads and an anode electrode. The detectable bacterial spores are at a linear range from 5 x 10(3) to 5 x 10(5) colony forming units (cfu) of Bacillus subtilis var. niger spores, 10(2) to 10(4) cfu of Bacillus anthrax spores and 10(2) to 10(6) cells of Escherichia coli O157:H7 in ECLIA. The unique MPFIA technique employs antibody-coated magnetic beads as solid phase in suspension for bacterial capture and concentration in a 96-well microplate format. Primary capturing antibodies, bacteria form a sandwich with alkaline phosphatase (AP)-labeled antibodies as reporter followed by a reaction with the AP substrate, AttoPhos(TM) to generate fluorescence for detection. Immunomagnetic separation permits direct isolating and concentrating bacterial cells from the crude samples, such as blood and environmental water. The results of MPFIA for detecting bacteria showed less sensitivity compared with that of ECLIA, however it provides a means for direct, high throughput screening bacteria from crude biological samples. Both ECLIA and MPFIA are rapid (less than one hour) and easy to use. (C) 1998 Elsevier Science B.V. All rights reserved.