[Laser-time-resolved fluorescence spectroscopy in immunoassays].
This paper described a laser-excited time-resolved fluoroimmunoassay set. It made lanthanide ion to couple the anhydrde of diethylenetriaminepentaacetic acid (DTPAA) for labeling antibodies. The experiment used polystyrene tap coated with HCV antigen as the solid phase and a chelate of the rare earth metal europium as fluorescent label. A nitrogen laser beam was used to excite the Eu3- chelates and after 60 microseconds delay time, the emission fluorescence was measured. Background fluorescence of short lifetimes caused by serum components and Raman scattering can be eliminated by set the delay time. In the system condition, fluorescent spectra and fluorescent lifetimes of Eu3+ beta-naphthoyltrifluroacetone (NTA) chelates were measured. The fluorescent lifetime value is 650 microseconds. The maximum emission wavelength is 613 nm. The linear range of europium ion concentration is 1 x 10(-7)-1 x 10(-11) g.mL-1 and the detection limit is 1 x 10(-13) g.mL-1. The relative standard deviation of determination (n = 12) for samples at 0.01 ng.mL-1 magnitude is 6.4%. Laser-TRFIA was also found to be suitable for diagnosis of HCV. The sensitivity and specificity were comparable to enzyme immunoassay. The result was obtained with laser-TRFIA for 29 human correlated well with enzyme immunoassay.
Pan
L
L
National Analytical Research Center of Eletrochemistry and Spectroscopy, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, 130022 Changchun.
Du
J
J
Xie
W
W
Du
Q
Q
Yun
Q
Q
D000818:Animals; D005063:Europium; D016231:Fluorescence Polarization Immunoassay / Q000379:methods*; D018937:Hepatitis C Antibodies / Q000097:blood*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood*; D028581:Lanthanoid Series Elements; D007834:Lasers; D011817:Rabbits
D018937:Hepatitis C Antibodies; D007074:Immunoglobulin G; D028581:Lanthanoid Series Elements; D005063:Europium