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Preparation method of colloidal gold chromatography test paper for fast detecting nitrofuran metabolite

A nitrofuracil and detection test paper technology, applied in the detection field, can solve the problems of limited application of ELISA method, high price, complicated operation process, etc., and achieve the effects of easy promotion, strong timeliness, and broad market prospects.

Inactive Publication Date: 2008-12-10
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ELISA method requires a supporting microplate reader and supporting reagents, and the operation process is still relatively complicated. Moreover, the country is currently in the research and development stage, and imported foreign products are expensive, so the application of the ELISA method is greatly restricted.

Method used

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  • Preparation method of colloidal gold chromatography test paper for fast detecting nitrofuran metabolite
  • Preparation method of colloidal gold chromatography test paper for fast detecting nitrofuran metabolite

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Preparation of test strips for the detection of nitrofurazone metabolites.

[0019] Firstly, it is necessary to prepare a carrier protein coupled to nitrofurazone metabolite derivatives, which is used to prepare corresponding detection lines (T lines) and antibodies; and it is necessary to prepare gold-labeled antibodies to nitrofurazone metabolite derivatives, which are used to prepare corresponding gold-labeled antibody fiber cotton; In addition, it is necessary to prepare goat anti-rabbit IgG antibody for the preparation of the control line (C line).

[0020] 1. Coupling of SEM derivatives to carrier proteins

[0021] Active ester method: Coupling SEM p-aldehyde benzoic acid derivative (CPSEM) with carrier protein BSA to prepare artificially bound antigen CPSEM-BSA. The specific preparation method is as follows:

[0022] (1) Take CPSEM 20.7mg (0.1mmol), N-hydroxysuccinimide NHS 17.25mg (0.15mmol), dicyclohexylcarbodiimide DCC 30.9mg (0.15mmol) dissolved ...

Embodiment 2

[0044] Embodiment 2, preparation detection test strip. Spray the coated antigen with a concentration of 1 mg / mL and goat anti-rabbit IgG on the coated film as the test line (T) and control line (C) respectively, and dry in an oven at 37°C for 10 minutes. In the same way, gold-labeled antibody with a concentration of 0.2 mg / mL was coated on the conjugate pad. The test strip is composed of a backing plate, on which a sample pad, a colloidal gold bonding pad, a coating film and a water-absorbing pad are glued in sequence. Cut the affixed board into 3mm wide strips, and then put the test strips together with the desiccant into an aluminum foil bag and seal them for storage.

Embodiment 3

[0045] Embodiment 3, the detection method of test strip.

[0046] Pretreatment of test samples: take 1 g of pork tissue homogenate, add 5 mL of distilled water, 0.5 mL of 1M HCl and 100 μL of 0.01 M benzaldehyde (ethanol solution), shake fully, and place at 60° C. overnight. Then add 1M of K 2 HPO 4 5 mL, 0.4 mL of 1M NaOH and 5 mL of ethyl acetate, shake vigorously for 30 seconds. Centrifuge at 4000 rpm for 10 minutes at room temperature, then transfer 2.5 mL of the ethyl acetate supernatant to a new container. Blow dry it with nitrogen at 45° C., redissolve it with n-hexane and mix with 1 mL of 0.01 MPBST (pH 7.4, 0.05% Tween-20). Centrifuge at 4000 rpm for 10 minutes at room temperature, and finally take the bottom aqueous phase for determination.

[0047] Detection operation method: insert the sample end of the SEM colloidal gold chromatography test strip into the sample solution to be tested, the insertion depth does not exceed the marked line, take out the test strip...

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Abstract

The invention relates to a preparation method of a colloid gold chromatography test strip for rapidly detecting furacilin metabolites, belonging to the detection technology field. The strip comprises a liner and a sample pad, a gold labeled binding pad, a packed capsule and a water absorbing pad which are connected successively on the liner. The gold labeled binding pad is made from glass fiber cotton for absorbing SEM derivative gold labeled antibodies of the furacilin metabolites. A detecting line (T) which is printed by the solution of SEM derivative coupling vector protein and a control line (C) which is printed by goat anti-rabbit IgG solution are arranged on the packed capsule. The two lines are parallel arranged. The colloid gold chromatography test strip is prepared based on polyclonal antibodies with high affinity which are labeled by colloid gold. The test strip has the advantages of strong specificity, high sensitivity, convenience, rapid speed, strong timeliness and onsiteoperation. The lowest detected amount can reach to 5ppb. After the test strip is added in a sample solution to be detected, the detecting result can be judged within 15 minutes. The preparation method of the colloid gold chromatography test strip for rapidly detecting the furacilin metabolites has the advantages of visual, directviewing and accurate result display, cost saving performance, wide application range and convenient popularization.

Description

technical field [0001] The invention discloses a method for preparing a colloidal gold chromatography test strip for rapid detection of nitrofurazone metabolites, belonging to the technical field of detection. Background technique [0002] Nitrofuran antibiotics mainly refer to Furazolidone, Nitrofurazone, Furantoin and Furaltadone, which are a class of broad-spectrum antibiotics that are effective against both Gram-positive and Gram-negative bacteria. Certain antibacterial effect. Furantoin is a synthetic nitrofuran broad-spectrum antibacterial drug, which can inhibit or kill a variety of Gram-positive bacteria and Gram-negative bacteria, as well as some protozoa and fungi. It is stable and has been widely used in livestock and aquaculture. Nitrofuran drugs and their corresponding metabolites include furazolidone (AOZ), furaltadone (AMOZ), nitrofuran (SEM) and nitrofurantoin (AHD). Nitrofuran drugs are metabolized quickly in animals, and the metabolites combined in tissu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52G01N33/558
Inventor 胥传来徐一平刘丽强
Owner JIANGNAN UNIV
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