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Detection kit of sulfamonomethoxine as well as preparation method and application thereof

A hexamethoxine and detection kit technology, applied in the field of biochemistry, can solve the problems of insufficient sensitivity of ELISA kits, unsuitability for rapid detection of samples, high cost of HPLC method, etc. Pre-processing simple effects

Inactive Publication Date: 2011-09-07
ZHENGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Commonly used detection methods for SMM at home and abroad include HPLC method and ELISA kit. The HPLC method is expensive and the pretreatment process is cumbersome, which is not suitable for the rapid detection of a large number of samples; the sensitivity of the ELISA kit is relatively low, and the detection limit is low (generally to 10 -9 g / ml), which cannot meet the requirements for the content determination of certain low-concentration substances

Method used

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  • Detection kit of sulfamonomethoxine as well as preparation method and application thereof
  • Detection kit of sulfamonomethoxine as well as preparation method and application thereof
  • Detection kit of sulfamonomethoxine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Sulfamethazine Enhanced Chemiluminescence ELISA Kit

[0041] The sulfamethazine detection kit of the present invention comprises 1 piece of 96-well microwell plate coated with anti-sulfamethazine polyclonal antibody, 1 bottle of HRP-SMM enzyme-labeled antigen, 5 bottles of SMM standard solution gradient solution (concentrations are respectively 0.6pg / ml, 1pg / ml, 6pg / ml, 60pg / ml, 100pg / ml), HRP-SMM sample diluent (0.05mol / L carbonate (CB) buffer), chemiluminescence base solution A, chemiluminescence base solution B. One bottle of washing solution (pH7.2 0.01mol / L phosphate (PBS) solution containing 0.05% Tween-20) and one instruction manual. Chemiluminescence base solution A is 2×10 -5 ~1×10 -3 mol / L luminol solution, 5×10 -5 ~1×10 -3 A mixed solution of mol / L p-hydroxyphenol solution and 0.1mol / L Tris-HCl at pH 8.0; chemiluminescence base solution B is 2×10 -5 ~1×10 -3 mol / L hydrogen peroxide solution.

Embodiment 2

[0042] Example 2 Preparation of Sulfamethazine Enhanced Chemiluminescent ELISA Kit

[0043] (1) Preparation of HRP-SMM sample diluent and washing solution

[0044] A. Prepare HRP-SMM sample dilution solution:

[0045] Sodium carbonate 1.59g

[0046] Sodium bicarbonate 2.93g

[0047] Add pure water to 1000ml, sterilize by filtration, and store at 4°C.

[0048] B. Preparation of washing liquid:

[0049] Tween-20 500

[0050] Add 0.01mol / L PBS (pH7.2) to 1000ml, sterilize by filtration, and store at 4°C.

[0051] 0.01mol / L PBS (pH7.2) formula:

[0052] 0.2mol / L disodium hydrogen phosphate solution (take Na 2 HPO 4 .12H 2 O 71.6g, add pure water to 1000ml)

[0053] 0.2mol / L sodium dihydrogen phosphate solution (take NaH 2 PO 4 .2H 2 O 35.6g, add pure water to 1000ml)

[0054] Take 0.2mol / L Na 2 HPO 4 Solution 80ml, 0.2mol / L NaH 2 PO 4 Solution 20ml and 17g NaCl, add pure water to 2000ml, filter and sterilize, and store at 4°C.

[0055] (2) Preparation of micr...

Embodiment 3

[0064] Embodiment 3 The application of test kit of the present invention and assay operating procedure

[0065] (1) Equilibration: Take out the sample and kit from the refrigerated environment, and equilibrate at 18~25°C for 30±5min

[0066] (2) Adding samples and enzyme-labeled antigen: Take a 96-well chemiluminescent microplate, preset a blank control well, and add 50 HRP-SMM sample diluent, add 50 Sample to be tested; then add 100 to all wells on the microplate HRP-SMM enzyme-labeled antigen, shake for 5 minutes, and incubate at 37°C for 1 hour.

[0067] (3) Plate washing: Shake off the liquid in the wells of the plate, rinse at least 4 times with washing solution, and pat dry on absorbent paper.

[0068] (4) Add luminescent substrate: Add 50 chemiluminescence base solution A and 50 chemiluminescence base solution B to each well successively , protected from light and shaken for 2 minutes.

[0069] (5) Determination: Measure the relative luminescence value (RLU) of...

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Abstract

The invention discloses a detection kit of sulfamonomethoxine as well as a preparation method and application thereof, belonging to the field of biochemistry. The detection kit comprises a micropore plate and a detection reagent, wherein the micropore plate is an opaque plastic plate, and each pore of the micropore plate is coated with a peridium antibody of sulfamonomethoxine with the concentration of 2mug / mL; and the detection reagent comprises an enzyme labeled antigen (HRP-SMM (Horseradish Peroxide- Sulfamonomethoxine)), a sulfamonomethoxine standard substance gradient solution, an enzyme labeled antigen sample diluent, a chemiluminescent base solution A, a chemiluminescent base solution B and a cleaning solution. The detection kit of the sulfamonomethoxine is used for detecting the sulfamonomethoxine content in milk, eggs, meats and products thereof; and the design method of the kit is novel, convenient for quick on-the-spot inspection on a large scale, has the advantages of low cost, simplicity for operation and high sensitivity and specificity and has better application prospect for detecting the sulfamonomethoxine residue of a veterinary medicine.

Description

technical field [0001] The invention relates to a drug residue detection kit, in particular to a veterinary drug sulfamethoxine detection kit, a preparation method and a detection method thereof, and belongs to the field of biochemistry. Background technique [0002] Sulfamethazine (SMM) has strong antibacterial and protozoan infection effects, and is widely used in the treatment and prevention of livestock and poultry diseases, and is widely used in livestock and poultry production as a feed additive due to its growth-promoting effect. application. Therefore, it is one of the veterinary drugs with the most serious residue problems, and its allergies, drug-resistant strains and other problems have attracted widespread attention. Therefore, since 2005, the Ministry of Agriculture has taken the residues of sulfa drugs in animal products as another key point to monitor after clenbuterol hydrochloride. When people eat animal food containing such drugs for a long time, the low-...

Claims

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Application Information

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IPC IPC(8): G01N33/543
Inventor 吴拥军屈凌波石杰
Owner ZHENGZHOU UNIV
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