Development of a homogeneous immunoassay based on the AlphaLISA method for the detection of chloramphenicol in milk, honey and eggs.
ABSTRACT A homogenous light-induced chemiluminescence immunoassay was developed using AlphaLISA technology for the detection of chloramphenicol (CAP). This technology is based on two different kinds of bead, namely light-sensitive donor beads and beads containing chemiluminescers, also called acceptor beads. A competitive CAP AlphaLISA method was established using artificial antigen-coated acceptor beads, polyclonal antibodies, biotinylated goat anti-rabbit IgG and streptavidin-coated donor beads.
The sensitivity of detection was 0.0086 ng mL⁻¹ and the working range was from 0.0096 to 25 ng mL⁻¹. The intra- and inter-assay coefficients of variation were both below 10%. The average recovery rates at spiked levels of 0.05-10 ng mL⁻¹ were 103.2, 108.4 and 91.6% for milk, honey and eggs respectively. The data obtained from the samples showed good correlation with ELISA results.
The CAP AlphaLISA method is highly sensitive, specific and rapid and is suitable for screening large quantities of samples.