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Furaltadone metabolite detection kit

A technology for detecting kits and furaltadone, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems that the accuracy, sensitivity, and specificity cannot be better than imported kits, and achieve the effect of good performance and convenient use

Inactive Publication Date: 2009-09-09
深圳市绿诗源生物技术有限公司
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] ELISA kits for the detection of furaltadone have been developed at home and abroad, but most of the domestic demand is imported from abroad, and domestically produced kits are not superior to imported reagents in terms of accuracy, sensitivity, and specificity. box

Method used

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  • Furaltadone metabolite detection kit

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Experimental program
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Effect test

preparation example Construction

[0048] d. Preparation and purification of monoclonal antibody: Using in vivo induction method, Balb / c mice (8 weeks old) were intraperitoneally injected with 0.5ml of sterilized paraffin oil per mouse, and 5×10 hybridoma cells were injected intraperitoneally after 7-14 days 5 -10 6 Ascites collected 7-10 days later, purified by octanoic acid-saturated ammonium sulfate method, divided into small bottles, and stored at -20°C;

[0049] e. Antibody freeze-dried powder can dry the ascites at 37°C and store it at -20°C;

[0050] f. Antibody working solution is to dilute the antibody at a concentration of 0.02-0.08 μg / ml with antibody diluent.

[0051] (2) The steps for preparing the polyclonal antibody of furaltadone metabolite derivatives are:

[0052] New Zealand white rabbits were adopted as immunized animals, and the immunogen (the conjugate of furaltadone metabolite derivative hapten and keyhole limpet hemocyanin) was immunized at 1 mg / kg. Mix complete emulsifier with Freund...

Embodiment 1

[0112] Example 1 Preparation of ELISA kit components for detecting furaltadone metabolites

[0113] 1. Antigen Synthesis

[0114] a. Synthesis of Coating Progen

[0115] The furaltadone metabolite is synthesized by the o-nitrobenzaldehyde derivative method to synthesize the furaltadone metabolite derivative hapten, and then the hapten is obtained by coupling the hapten with the bovine gamma globulin carrier protein by the active ester method through a diazotization reaction.

[0116] b. Synthesis of Immunogen

[0117] The furaltadone metabolite was synthesized by the o-nitrobenzaldehyde derivative method to synthesize the furaltadone metabolite derivative hapten, and then the hapten was coupled with the keyhole limpet hemocyanin carrier protein by the active ester method through the diazotization reaction. Get it.

[0118] 2. Preparation of mouse monoclonal antibody against furaltadone metabolite derivatives

[0119] a. Animal immunity

[0120] use. Balb / c mice were used...

Embodiment 2

[0131] Example 2 The formation of an enzyme-linked immunosorbent assay kit for detecting furaltadone metabolites

[0132] An enzyme-linked immunosorbent assay kit for detecting furaltadone metabolites was constructed to include the following components:

[0133] (1) an enzyme-linked plate coated with furaltadone metabolite derivative antigen;

[0134] (2) specific mouse antibody of furaltadone metabolite derivatives labeled with horseradish peroxidase;

[0135] (3) 6 bottles of furaltadone metabolite standard solution, the concentrations were 0μg / L, 0.05μg / L, 0.15μg / L, 0.45μg / L, 1.35g / L, 4.05μg / L;

[0136] (4) Substrate chromogenic solution hydrogen peroxide and tetramethylbenzidine sulfate mixed solution;

[0137] (5) The stop solution is 2mol / L sulfuric acid buffer;

[0138] (6) The concentrated washing liquid is deionized water or ultrapure water;

[0139] (7) Antibody diluent is pH 7.4, 0.08mol / L phosphate buffer containing 0.3% gelatin, 5‰ Tween-80 and 5‰ methanol.

...

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Abstract

The invention relates to a furaltadone metabolite detection kit and provides an elisa kit for detecting furaltadone metabolite, comprising an enzyme yoke plate with an encrusting furaltadone metabolite derivate antigen, an enzyme yoke plate with an enzyme-labeling furaltadone metabolite derivate specific antibody or an encrusting furaltadone metabolite derivate specific antibody, and an enzyme-labeling furaltadone metabolite derivate antigen, wherein the furaltadone metabolite derivate antigen is obtained by coupling a furaltadone metabolite derivate hapten and carrier protein by an active ester method. The kit is convenient to use and has favorable performance in aspects, such as accuracy, sensitivity, specificity, and the like.

Description

technical field [0001] The invention relates to a kit for detecting furaltadone metabolites. Background technique [0002] Nitrofuran drugs are synthetic broad-spectrum antibiotics, which have very good antibacterial effects and pharmacokinetic properties, and have been widely used as growth-promoting additives for livestock, poultry, and artificially farmed aquatic products. However, through long-term laboratory research, it has been found that both nitrofuran drugs and metabolites can cause cancer and gene mutations in experimental animals, and it can be inferred that humans have long-term use of such drugs or long-term consumption of livestock supplemented with such growth-promoting agents , poultry, and artificially farmed aquatic products can also undergo canceration and gene mutations. Therefore, such drugs are prohibited from being used in treatment and feed. [0003] In 2002, my country promulgated a ban on the use of nitrofuran antibiotics. In the "List of Veterin...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/545G01N33/577
Inventor 聂继斌唐俊杨宏温俊梅齐欣吴青李成杨宗繁
Owner 深圳市绿诗源生物技术有限公司
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