Abstract
chemiluminescent immunoassays (CLIA) based on gold magnetic nanoparticles (Au-MNPs) were developed for rapid analysis of chloramphen icol (CAP) in milk sample. Anti-CAP antibodies were immobilized on the surfaces of Au-MNPs, luminol (Method I) and 2′,6′-DiMethylcarbonylphenyl-10-sulfopropylacridinium-9-carboxylate 4′-NHS Ester, (NSP-DMAE-NHS, Method II) were exploited in competitive CLIA for CAP detection in milk using a homemade luminescent measurement system. The sensitivities and limits of detection (LODs) of the two methods were obtained according to the inhibition curves. It indicated that NSP-DMAE-NHS as luminescence reagent (reaction II) was more sensitive and effective than luminol (reaction I). The LOD of reaction II reached 0.008 ng/ml while it was 4 ng/ml in reaction I. Moreover, the linear range of the inhibition curve of the former was wider than that of the latter. Such results indicated that the proposed CLIA stratery employing NSP-DMAE-NHS was more sensitive than other immunoassay method for CAP detection.
Highlights
Chloramphenicol (CAP), a kind of cheap and effective bacteriostatic broad-spectrum antibiotics, was widely used in animal husbandry to treat kinds of sensitive bacteria infection
It was found that CAP had serious adverse effects against human bodies, leading to bone marrow depression and even aplastic anemia [1,2]. Due to these side effects, it was banned from use in animal feeds and food in the European Union (EU), Switzerland, USA and other countries [3,4]
This paper explored the Au-MNPs-based chemiluminescent immunoassays (CLIA) methods for CAP detection in milk samples respectively with luminol and NSP-DMAE-NHS as luminescence reagent
Summary
Chloramphenicol (CAP), a kind of cheap and effective bacteriostatic broad-spectrum antibiotics, was widely used in animal husbandry to treat kinds of sensitive bacteria infection. RIAs have high sensitivity but exists radioactive waste, and ELISAs have shortcoming of higher limit of detection (LOD). Chemiluminescent immunoassays (CLIAs) have some features such as non-radioactive wastes, simple instrumentation, the very low limit of detection (LOD), high sensitivity, wide dynamic range and rapid analysis time in only several seconds. Those characteristics make it broadly used in clinical, environment, food and factory analysis in recent years [14,15]. This paper explored the Au-MNPs-based CLIA methods for CAP detection in milk samples respectively with luminol (reaction I) and NSP-DMAE-NHS (reaction II) as luminescence reagent
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