Light-induced chemiluminescent immunoassay kit and test method for chloramphenicol
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A light-induced chemiluminescence and immunoassay technology, which is applied in material excitation analysis, fluorescence/phosphorescence, biological testing, etc., can solve problems such as adverse effects, bacterial flora imbalance, and allergic reactions, and achieve short detection time, easy operation, and high sensitivity. high effect
Inactive Publication Date: 2009-12-16
JIANGSU INST OF NUCLEAR MEDICINE
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In mild cases, the balance of normal flora in the human body will be destroyed, and the flora will be out of balance, causing the human body to produce drug-resistant bacteria beads, which will bring adverse effects on the use of antibiotics for future illnesses; people with antibiotic allergies will have allergic reactions, endangering health
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Embodiment 1
[0021] Embodiment 1 preparation kit
[0022] Preparation of luminescent particles coated with CAP-OVA:
[0023] Add 1 mg of luminescent particles to a centrifuge tube, add 12.5 μL 1% Tween-20, 0.05 mg CAP-OVA artificial antigen, 10 μL sodium borohydride, and use 0.1 M, pH 6.0 2-(N-morpholine)ethanesulfonic acid ( The volume of MES) buffer was added to 200 μL, and the reaction was shaken at 37°C for 48 hours in the dark. Add 10 μL of 0.3M, pH 5.0 carboxymethoxylamine hemihydrochloride (CMO) solution to block unbound sites, incubate at 37°C in the dark for 1 hour, and then centrifuge to separate the luminescent particles coated with CAP-OVA. Ready to use after dilution.
[0024] Preparation of CAP standard reagents: (0ng / mL, 0.02ng / mL, 0.1ng / mL, 1ng / mL, 10ng / mL, 25ng / mL), obtained by dilution from pure CAP, diluent containing 10% methanol PBS (0.05mmol / L, pH 7.4).
[0025] The composition of the kit:
[0026] (1) White opaque microwell plate (12 strips x 8 wells, which can ...
[0038] Honey sample processing: Mix 2g sample with 4mL distilled water, add 4mL ethyl acetate, stopper and shake for 10min, centrifuge at 3000g for 10min. Take 2mL of the supernatant into a clean glass tube, evaporate to dryness under gentle nitrogen flow at 50°C, and fully dissolve the residue in 1mL of standard diluent for later use.
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Abstract
The invention discloses a light-induced chemiluminescent immunoassay kit and a test method for chloramphenicol (CAP) and belongs to the technical field of light-induced chemiluminescent immunoassay technology. A nontransparent white microporous plate is sequentially added with CAP-OVA coated luminous particles, a CAP standard substance or a sample to be tested, a rabbit anti-CAP antibody and a biotin goat anti-rabbit antibody for reaction without light and then is added with streptavidin-coated light sensitive particles for after-incubation test. The CAP-OVA on the luminous particles and free CAP compete to be connected to the CAP antibody to form a compound body with the biotin goat anti-rabbit antibody and the streptavidin-coated light sensitive particles. Under the excitation of red light, the compound body transfers energy to the luminous particles through the generation and transmission of singlet ionized oxygen for generating fluorescence. A light induced chemiluminescent detector is used to detect the intensity of an optical signal, and the CAP content of the sample can be determined by referring to a standard curve according on the basis that the intensity of the optical signal is in inverse proportion to the CAP concentration of the sample. The method is used for determining the CAP content of foods such as honey, milk and eggs. The kit is simple in structure, short in determination time, high in sensitivity and simple and convenient in operation.
Description
technical field [0001] A light-induced chemiluminescence immunoassay kit for chloramphenicol and a detection method thereof belong to the technical field of light-induced chemiluminescence immunoassay (LICLIA), and are used for detecting CAP content in food. Background technique [0002] Chloramphenicol (Chloramphenicol, CAP) is a cheap and efficient broad-spectrum antibiotic, which has a good inhibitory effect on both Gram-positive and Gram-negative bacteria, so it was once widely used in agriculture and animal husbandry. However, animal-derived foods are ingested by the human body for a long time along the food chain, which can cause many diseases. In mild cases, the balance of normal flora in the human body will be destroyed, and flora imbalance will cause the human body to produce drug-resistant bacteria beads, which will bring adverse effects on the use of antibiotics for future illnesses; people with antibiotic allergies will have allergic reactions, endangering health...
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