H. Yu et al., Detection of biological threat agents by immunomagnetic microsphere-based solid phase fluorogenic- and electro-chemiluminescence, BIOSENS BIO, 14(10-11), 2000, pp. 829-840
This article reviews the recent development of two solid-phase chemilumines
cence-based techniques, fluorogenic-chemiluminescence (FCL) and electro-che
miluminescence (ECL) for detection of biological threat agents. Both techni
ques entail a labeled sandwich immunoassay. The objectives of this work are
to develop advanced techniques for sensitive and effective detection of a
target analyte, particularly in cases where the analysis includes complex s
amples containing multiple contaminating factors. Other important considera
tions in developing such detection techniques include the ease of use, the
rapid determination of the results, and system automation for field applica
tions. In FCL, alkaline phosphatase is used as a label and this technique u
tilizes the dual features of fluorescence and visual color generated upon t
he presence of the fluorogenic compound, AttoPhos(TM). The assay reaction i
s determined by measuring the fluorescence. In ECL, the label is a rutheniu
m-trisbipyridal, which is excited to a higher energy state by an electric c
urrent-driven redox reaction, and the extent of the reaction is assessed vi
a photon emission. Both techniques depend upon the magnetic separation tech
nique as a means to isolate the target immunological agents from the sample
for analysis. This magnetic capture system allows for a reaction to occur
on the electron effective-transfer zone in the ECL and also provides the re
action site for the labeled sandwich in the FCL. Comparative studies of the
se two techniques for detection of biological threat agents have been perfo
rmed and the advantages of using magnetic microspheres versus conventional
solid-phase matrices are discussed. (C) 2000 Elsevier Science S.A. All righ
ts reserved.