Nicarbazin detection kit and application thereof

A detection kit, the technology of nicarbazine, which is applied in the field of biotechnology detection, can solve the problems of long time required for detection reaction, low detection sensitivity, complicated detection steps, etc., and achieves simple structure, good portability and low detection cost. Effect

Active Publication Date: 2021-03-09
北京元恩生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invention patent application publication number CN106771137 discloses an enzyme-linked immunosorbent assay kit for detecting nicarbazine and its application, which discloses an enzyme-linked immunosorbent assay kit, which has an enzyme-labeled plate coated with a nicarbazine-coupled antigen, nicarbazine Nicarbazine monoclonal antibody, enzyme-labeled anti-antibody, nicarbazine standard solution, substrate chromogenic solution, stop solution, the detection steps of the prior art ELISA kit are still relatively cumbersome, and the time required for the detection reaction is longer, and Detection sensitivity is still not high

Method used

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  • Nicarbazin detection kit and application thereof
  • Nicarbazin detection kit and application thereof
  • Nicarbazin detection kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1. Nicarbazine detection kit contains components:

[0038] (1) A 96-well microtiter plate coated with a nicarbazine hapten-OVA conjugate;

[0039] (2) Enzyme-labeled antibody working solution, specifically horseradish peroxidase-labeled nicarbazine monoclonal antibody, 7 mL / bottle. 8% glycerol, 0.1% EDTA, 5% trehalose, and 0.01% Proclin300 were added to the enzyme-labeled antibody working solution as antibody stabilizers, and the percentages are mass percentages.

[0040] (3) Nicarbazine marker standard 0μg / L, 0.025μg / L, 0.075μg / L, 0.225μg / L, 0.675μg / L, 2.025μg / L, 1mL / bottle.

[0041] (4) 20×concentrated washing solution, containing 1% polyethylene glycol 400 monooleate, 0.1mol / L MES buffer, pH6.0, 30mL / bottle, use deionized water at a ratio of 1:19 The ratio is diluted.

[0042] (5) 20×concentrated complex solution, the composition is 50% DMF, 0.01mol / L carbonate buffer, pH 9.0, 10mL / bottle, diluted with methanol at a ratio of 1:19.

[0043] (6) The substrate A sol...

Embodiment 2

[0086] The detection of nicarbazine marker in the chicken sample of embodiment 2

[0087] 1. Chicken sample pretreatment method

[0088] Weigh 1±0.05g sample into a 10mL centrifuge tube, add 3mL of absolute ethanol, vortex for 5min; centrifuge at 5000rpm room temperature for 5min; take 25μL of the clear liquid in the upper layer, add 975μL of reconstitution working solution, shake and mix for 30s; take 50μL for analysis .

[0089] 2. Standard curve fitting

[0090] The standard curve is composed of 6 concentrations (0μg / L, 0.025μg / L, 0.075μg / L, 0.225μg / L, 0.675μg / L, 2.025μg / L), and each of the 6 standard points is determined twice, with Determine the working concentration range. Taking the logarithmic value of the nicarbazine marker standard substance concentration as the abscissa, the inhibition rate (B / B) of each corresponding concentration 0 %) is the ordinate, and a standard curve is made. see results figure 2 .

[0091] 3. Sample precision and accuracy test

[0092...

Embodiment 3

[0096] (1) A 96-well microtiter plate coated with a nicarbazine hapten-HSA conjugate;

[0097] (2) Enzyme-labeled antibody working solution, specifically horseradish peroxidase-labeled nicarbazine polyclonal antibody, 7 mL / bottle.

[0098] (3) Nicarbazine marker standard 0μg / L, 0.025μg / L, 0.075μg / L, 0.225μg / L, 0.675μg / L, 2.025μg / L, 1mL / bottle.

[0099] (4) 20×concentrated washing solution, the composition is 1% polyethylene glycol 400 monooleate, 0.1mol / L MES buffer solution, pH 6.0, 30mL / bottle.

[0100] (5) 20×concentrated complex solution, the composition is 50% DMF, 0.01mol / L carbonate buffer solution, pH 9.0, 10mL / bottle.

[0101] (6) The substrate A solution is hydrogen peroxide, and the substrate B solution is TMB, both 7mL / bottle.

[0102] (7) The stop solution is 2mol / L H 2 SO 4 , 7mL / bottle.

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Abstract

The invention provides a nicarbazin detection kit. The nicarbazin detection kit comprises a 96-hole elisa plate, an enzyme marker, a nicarbazin marker standard substance, a 20*concentrated washing solution, a 20*concentrated reconstitution fluid, a substrate solution A, a substrate solution B and a stop solution, wherein the 96-hole elisa plate is coated with a nicarbazin hapten carrier protein, the enzyme marker is a nicarbazin specific antibody marked by horse radish peroxidase. The invention also provides an application of the nicarbazin detection kit in detection of chicken samples. The nicarbazin detection kit adopts a one-step method to detect the nicarbazin marker, is high in detection sensitivity, good in accuracy and precision of a detection result, simple in structure, convenientto use, good in portability, low in requirement on detection personnel and suitable for rapid detection of large-batch samples on site.

Description

technical field [0001] The invention belongs to the field of biotechnology detection, and in particular relates to a nicarbazine ELISA rapid detection kit and its application. Background technique [0002] Nicarbazine, also known as cochowerine and dinitrourea dipyrimidinol, is a 1:1 complex composed of dinitrodiphenylurea (DNC) and hydroxydimethylpyrimidine (HDP). Spectrum, high efficiency, stable performance feed additive. Nicarbazine for the prevention of coccidiosis in chickens and turkeys. Although nicarbazine is a compound composed of DNC and HDP, because the HDP component in the compound can be quickly excreted through urine in animals, and the anticoccidial active component DNC ​​is slowly excreted through feces, so the international Dinitrodiphenylcarbamide (DNC) is stipulated as the marker of nicarbazine residues above. [0003] At present, nicarbazine is widely used, but it will cause different degrees of residues in poultry muscles and other tissues after feed...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/543G01N33/58
CPCG01N33/5308G01N33/581G01N33/543
Inventor 王世恩郝士元杨昌松
Owner 北京元恩生物技术有限公司
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