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

A technology for the detection of nitrofurantoin metabolites and test strips, applied in the detection field, can solve the problems of complex operation process, high price, and application limitation of ELISA method, and achieve the effect of broad market prospect, strong timeliness, and easy promotion

Inactive Publication Date: 2008-12-10
JIANGNAN UNIV
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  • 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 c

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0019] First of all, it is necessary to prepare a carrier protein coupled to derivatives of nitrofurantoin metabolites, which is used to prepare corresponding detection lines (T lines) and antibodies; and it is necessary to prepare gold-labeled antibodies to derivatives of nitrofurantoin metabolites, which is 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 AHD derivatives with carrier protein

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

[0022] (1) Take CPAHD 24.7mg (0.1mmol), N-hydroxysuccinimide NHS 17.25mg (0.15mmol), dicyclohexylcarbodiimide DCC 30.9mg (0...

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 AHD 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 colloidal gold chromatography test paper tape which can test nitrofurantoin metabolite and belongs to the field of testing technology. The test paper tape consists of a lining board and a sample gasket, a gold standard bonding gasket, peridium membrane and an absorbent gasket which are jointed together in sequence on the lining board. The gold standard bonding gasket is glass fiber cotton which can absorb the gold-standard antibody of AHD derivative of nitrofurantoin metabolite. A test line (T) printed by the coupling carrier protein solution of AHD derivative and a control line (C) printed by Goat anti-Rb IgG solution are arranged in parallel with each other on the peridium membrane. The colloidal gold chromatography test paper tape is prepared on the bases that colloidal gold is used for marking a high-affinity polyclonal antibody. The test paper tape has high specificity and sensitivity, and the minimum limit which can be tested can reach 5ppb. The test paper tape can be used easily and conveniently on the spot without any other test reagent and instruments, and the test result can be obtained quickly, thereby achieving high timeliness. After the test paper tape is put into the tested sample solution, the test results can be determined in 15 minutes, and the results can be shown vividly, directly and correctly. The test paper tape can save cost and has wide scope of application; therefore, the test paper tape can be promoted conveniently.

Description

technical field [0001] The invention discloses a method for preparing a colloidal gold chromatography test strip for rapid detection of nitrofurantoin 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 has inhibitory or killing effects on 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...

Claims

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Application Information

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IPC IPC(8): G01N33/52G01N33/558
Inventor 胥传来徐一平刘丽强
Owner JIANGNAN UNIV
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