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Cistofuran metabolite detection reagent kit

A technology for nitrofurantoin metabolites and detection kits, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of accuracy, sensitivity, and specificity that cannot be better than imported kits, and achieves the effects of good performance and convenient use.

Inactive Publication Date: 2009-01-21
深圳市绿诗源生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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

Method used

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  • Cistofuran metabolite detection reagent kit
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  • Cistofuran metabolite detection reagent kit

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0047] d. Preparation and purification of monoclonal antibody: In vivo induction method, intraperitoneal injection of 0.5ml of sterilized paraffin oil into Balb / c mice (8 weeks old), 7-14 days later, intraperitoneal injection of hybridoma cells 5×10 5 ~10 6 One per mouse, collect ascites after 7-10 days, purify the ascites with octanoic acid-saturated ammonium sulfate method, divide into vials, and store at -20°C;

[0048] e. Antibody freeze-dried powder can dry the ascites at 37°C and store it at -20°C;

[0049] f. Antibody working solution is to dilute the antibody at a concentration of 0.02~0.08μg / ml with antibody diluent.

[0050] (2) The steps for the preparation of polyclonal antibodies against nitrofurantoin metabolite derivatives are:

[0051] New Zealand white rabbits were used as immunized animals, the immunogen (the conjugate of nitrofurantoin metabolite derivative hapten and keyhole limpet hemocyanin) was immunized at 1 mg / kg, and the immunogen was mixed with an ...

Embodiment 1

[0110] Example 1 Preparation of ELISA Kit Components for Detecting Nitrofurantoin Metabolites

[0111] 1. Antigen Synthesis

[0112] a. Synthesis of Coating Progen

[0113] The nitrofurantoin metabolite is synthesized by the derivative method to synthesize the nitrofurantoin 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 the diazotization reaction.

[0114] b. Synthesis of Immunogen

[0115] The nitrofurantoin metabolite is synthesized by the derivative method to synthesize the nitrofurantoin metabolite derivative hapten, and then the hapten is obtained by coupling the hapten with the keyhole limpet hemocyanin carrier protein by the active ester method through a diazotization reaction.

[0116] 2. Preparation of mouse monoclonal antibody against nitrofurantoin metabolite derivatives

[0117] a. Animal immunity

[0118] use. Balb / c mice were used as immun...

Embodiment 2

[0129] Example 2 Formation of ELISA Kit for Detecting Nitrofurantoin Metabolites

[0130] An enzyme-linked immunosorbent assay kit for detecting nitrofurantoin metabolites was constructed to include the following components:

[0131] (1) Enzyme-linked plates coated with nitrofurantoin metabolite derivative antigens;

[0132] (2) specific mouse antibody of nitrofurantoin metabolite derivatives labeled with horseradish peroxidase;

[0133] (4) 6 bottles of nitrofurantoin metabolite standard solution, the concentrations were 0 μg / L, 0.05 μg / L, 0.15 μg / L, 0.45 μg / L, 1.35 g / L, 4.05 μg / L;

[0134] (5) Substrate chromogenic solution hydrogen peroxide and tetramethylbenzidine sulfate mixed solution;

[0135] (6) The stop solution is 2mol / L sulfuric acid buffer;

[0136] (7) The concentrated washing liquid is deionized water or ultrapure water;

[0137] (8) Antibody diluent is pH 7.4, 0.08mol / L, phosphate buffer containing 0.3% gelatin, 5‰ Tween-80 and 5‰ methanol.

[0138] (9) Th...

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Abstract

The invention relates to a nitrofurantoin metabolite detection kit for detecting nitrofurantoin metabolites, which comprises an enzyme-linked plate which is coated with nitrofurantoin metabolite derivant antigen and an enzyme label nitrofurantoin metabolite specific antibody or an enzyme-linked plate which is coated with nitrofurantoin metabolite derivant specific antibody and an enzyme label nitrofurantoin metabolite antigen, wherein the nitrofurantoin metabolite derivant antigen is got through coupling nitrofurantoin metabolite derivant hapten and carrier protein through adopting an active ester method. The invention has the advantages of excellent property in the aspects of accuracy, sensitivity, specificity and other aspects.

Description

technical field [0001] The invention relates to a kit for detecting nitrofurantoin 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 Vete...

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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