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Colloidal gold test paper card for detecting quinolones medicament relict

A technology of colloidal gold test paper and quinolones, which is applied in the field of immunochemical rapid detection of veterinary drug residues, can solve the problems of being unsuitable for on-site monitoring and screening of a large number of samples, unfavorable for the promotion and use of grassroots units, and complex instruments and equipment, and achieves short detection time , low cost and high sensitivity

Active Publication Date: 2008-05-07
BEIJING WANGER BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One is chromatographic analysis, such as high performance liquid chromatography (HPLC), which is not suitable for on-site monitoring and screening of a large number of samples due to complex instruments and tedious processes
The other is an immunological method, such as enzyme-linked immunosorbent assay (ELISA), which has the disadvantages of long detection time and high cost, which is not conducive to popularization and use in grassroots units.
Moreover, both of these methods require professional and technical personnel to operate

Method used

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  • Colloidal gold test paper card for detecting quinolones medicament relict

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 Preparation of QNs detection test paper card

[0031] 1. Synthesis and identification of QNs hapten-carrier protein conjugates

[0032] (1) Preparation of QNs hapten

[0033] Take pipemidic acid (8-ethyl-5-oxo-5,8-dihydro-2-(1-piperazinyl)pyrido[2,3-d]pyrimidine-6-carboxylic acid) 500mg dissolved in In 30ml of water, add 200mg of EDC to react at room temperature for 1h, then add 150mg of p-aminophenylacetic acid to react overnight. Then the QNs hapten was extracted by acid washing, and dried to obtain 280 mg. The QNs hapten was 8-ethyl-5-oxo-5,8-dihydro-2-(1-piperazinyl)pyrido[2,3 -d] pyrimidine-6-amidophenylacetic acid.

[0034] (2) Synthesis of QNs hapten-human serum protein conjugate

[0035] Weigh 90 mg of human serum albumin (HSA), add 10 ml of PBS to dissolve, and then add 10 ml of N,N-dimethylamide (DMF) while stirring in an ice bath to obtain an HSA solution. Weigh 7.8 mg of the QNs hapten, dissolve it in 1.5 ml of DMF, add 8 μL of tributylamin...

Embodiment 2

[0063] The detection of QNs residue in the sample of embodiment 2

[0064] 1. Sample pretreatment

[0065] (1) Animal tissue pretreatment (chicken, chicken liver, pork, pork liver)

[0066] Weigh 2.0±0.05g of the homogenized tissue sample into a centrifuge tube, and tightly cap the bottle. Place the centrifuge tube containing the sample in a slightly boiling water bath for 10 minutes, absorb more than three drops of the boiled solution and pour it into a 1.5ml centrifuge tube. If there is obvious yellow turbidity, please centrifuge, and then use the supernatant as the sample solution to be tested.

[0067] (2) Pretreatment of egg samples

[0068] Homogenize the egg sample, weigh 2.0±0.05g into a centrifuge tube, and tightly cap the bottle. Place the centrifuge tube containing the sample in a slightly boiling water bath for 10 minutes, absorb more than three drops of the boiled solution and pour it into a 1.5ml centrifuge tube. Please use the supernatant as the sample solu...

Embodiment 3

[0077] Example 3 sample detection example

[0078] Take 15 samples of eggs, chicken, and honey with known QNs concentration greater than 10ng / g and 15 samples of each negative sample, repeat the test twice for each sample, and calculate the negative and positive rates. The results are shown in Table 1, Table 2 and Table 3.

[0079] Table 1 Negative and positive rate detection of egg samples

[0080] Negative

[0081] Table 2 Positive and negative rate detection of chicken samples

[0082]

[0083] result

[0084] Table 3 Negative and positive rate detection of honey samples

[0085]

[0086] The results show that there are 15 samples of eggs, chicken, and honey, and 15 samples of their own negative samples. Each sample is tested twice. The coincidence rate of negative and positive in eggs is 100%, and the coincidence rate of negative and positive in chicken is 100%. %. The false positive rate in honey is less than 7%, the positiv...

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Abstract

The invention provides an immune colloidal gold test paper card for testing quinolones, comprising a reaction film, a sample pad, a conjugate release pad, a water absorbing pad, and a back liner, wherein, the reaction film comprises a test zone coated with quinolones-carrier protein conjugate and a quality control zone coated with sheep-anti-mouse IgG; the conjugate release pad coats quinolones monoclonal antibody-colloidal gold marker. The invention also provides the method using the test paper card to test quinolones, which comprises the following steps: firstly, pre-treating sample; secondly, testing the sample with the test paper card; thirdly, analyzing test result. The invention can be used to test residual quantity of quinolones in animal derived food such as chicken, chicken liver, pork, pig liver, urine, and honey, and has the advantages of simple operation, high sensitivity, fast test, low cost, on-site supervision, and satisfaction of test requirement of a large quantity of samples.

Description

technical field [0001] The invention relates to the technical field of rapid immunochemical detection of veterinary drug residues, and more specifically relates to a colloidal gold test paper card for detecting quinolone drug residues, which can quickly detect quinolone drug residues by means of immunochromatographic reaction of colloidal gold labeling and color development. Background technique [0002] Quinolones (QNs) are a class of very important broad-spectrum antibiotics that have developed rapidly in the past 20 years. In terms of chemical structure, this class of drugs belongs to pyridonecarboxylic acids (PCAs), commonly known as "quinolones" (QNs). QNs inhibit bacterial DNA helicase, have a wide antibacterial spectrum, high efficiency, low toxicity, and strong tissue penetration ability, and have become one of the most important anti-infective drugs in veterinary clinics and aquaculture. Sex attracts a lot of attention. In tissues, the marked residues of enrofloxa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/532C12N5/12
Inventor 沈建忠何方洋万宇平冯才伟赵正苗吴小平冯才茂朱亮朱颜
Owner BEIJING WANGER BIOTECH
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