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Immune colloid gold test paper strip for detecting sulfadiazine residue and its preparation method

A technology of sulfadiazine and colloidal gold, which is applied in the field of immunochemical rapid detection of veterinary drug residues, can solve the problems of complicated operation process, high detection cost, difficult to popularize, etc. Effect

Inactive Publication Date: 2009-04-29
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Chromatographic technology is very effective, accurate and sensitive for the detection of sulfadiazine, but it also has the following disadvantages: (1) The pretreatment procedure of the sample to be tested is cumbersome and time-consuming (2) It needs professionally trained technicians to operate, and the operators must have rich experience Relevant experience, operators must understand the various interference factors that affect chromatographic analysis, understand the advantages and disadvantages of the pretreatment method used, in order to obtain reliable analysis results; popular among enterprises
(4) The requirements for instrument maintenance are high, and the quality of maintenance directly affects the accuracy of the analysis results; (5) The detection cost is high
The disadvantages are: (1) special instruments and equipment such as microplate reader are required to be used together, which is not conducive to popularization and use in grassroots units; (2) detection operators need to undergo professional training; (3) the operation process is relatively complicated, The time required for the detection is relatively long, and the whole process takes 2h-4h; (4) The cost of the detection is relatively high, and a single detection cannot be realized

Method used

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  • Immune colloid gold test paper strip for detecting sulfadiazine residue and its preparation method
  • Immune colloid gold test paper strip for detecting sulfadiazine residue and its preparation method
  • Immune colloid gold test paper strip for detecting sulfadiazine residue and its preparation method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 (preparation embodiment)

[0034] Preparation method of immunocolloidal gold test strip for detecting sulfadiazine

[0035] 1. Preparation of sulfadiazine-carrier protein conjugates

[0036] Synthesis of sulfadiazine-carrier protein conjugates:

[0037] (1) Accurately weigh 40mg of sulfadiazine and dissolve in 2ml of 1M NaOH and 1.1ml of 1% NaNO 2 The mixed solution of sulfadiazine was prepared.

[0038] (2) Under ice-water bath and magnetic stirring, add the sulfadiazine solution dropwise into 2.4ml 1mol / l HCl for diazotization (Itaru Y, KohjiI. Enzyme immunoassay for clenbuterol, an β 2 -adrnergicstimulant [J]. Journal of immunoassay, 1982, 3 (2): 155-171) reaction, continue to react for 5 minutes after adding.

[0039] (3) Under magnetic stirring, the diazotization products were added dropwise to human serum albumin (HSA), bovine serum albumin (BSA) and ovalbumin (OVA) solutions, and the pH value was adjusted to 9.5, 4°C and stirred slowly overnight...

Embodiment 2

[0060] Embodiment 2 (application embodiment)

[0061] Method for using the immunocolloidal gold test strip for detecting sulfadiazine

[0062] 1. Sample pretreatment

[0063] (1) Pretreatment of tissue samples

[0064] Homogenize the tissue sample to be tested, take 3g of the tissue sample to be tested, add 6ml of ethyl acetate, shake up and down for 10min, centrifuge at 2000g at room temperature for 10min, take 2ml of the upper liquid and evaporate it to dryness or dry it under nitrogen flow, and use 1ml of 0.01M phosphoric acid pH7.4 Salt buffer solution (recipe: NaCl 8g, KCl 0.2g, NaCl 2 HPO 4 12H 2 O 2.9g, KH 2 PO 4 0.2g, and distilled water to 1000ml) to dissolve the residue.

[0065] (2) Pretreatment of honey samples

[0066] Take 2g of honey sample and add 4ml of double distilled water to dissolve, add 4ml of ethyl acetate and shake up and down for 10min, centrifuge at 3000g at room temperature for 10min, take 1ml of the upper liquid and evaporate to dryness or b...

Embodiment 3

[0075] Embodiment 3 (application embodiment)

[0076] Application of immunocolloidal gold test strip for detection of sulfadiazine

[0077] 1. Specificity test

[0078] Prepare sulfadiazine, sulfadiazine sodium, sulfamethoxazole, sulfamethoxazole, sulfamethoxine, sulfaquinoxaline, sulfadimethoxine, sulfamethoxine, clenbuterol, and chloramphenicol into a sample with a concentration of 500ppb. Test according to the method described in Example 2. Test result (see table 1) shows, sulfadiazine, sulfadiazine sodium sample detection line has no color to appear, and red band appears on quality control line simultaneously, and sulfamethazine, sulfamethoxazole, sulfamethoxazole, sulfamethoxine, sulfamethoxazole The color of test strips of quinoxaline, sulfadimethoxine, sulfamethoxine, clenbuterol, and chloramphenicol samples is the same as that of standard test strips, and red bars appear on the quality control line band, which shows that sulfamethazine, sulfamethoxazole, sulfametho...

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Abstract

This invention discloses a way for fast checking the immunological colloid gold test strip left by the sulfadiazine and it's making method, which belongs to the immunochemistry fast measure technique filed. The test strip of this invention includes: the sample mat, the combining mat, the cellulose nitrate film, the sopping mat and the PVC backing, which characterized in that the sample mat, the combining mat, the cellulose nitrate film and the sopping mat inhibit the PVC backing in order; the combining mat is marked with the anti-sulfadiazine monoclonal antibody-colloid gold marker, and the monoclonal antibody is obtained by the secretion of the hybridomas cell line BDRXN(the conserving number is CCTCC-C200522) The cellulose nitrate film is covered with the checking line that is composed of the sulfadiazine-carrier protein joined matter and the quality control line that is composed of the rabbit-anti-mouse IgG. The test strip of this invention has the following advantages: the sensitivity is high, the special isomerism is strong, the operation is simple and checking is fast and exactly.

Description

technical field [0001] The invention belongs to the technical field of immunochemical rapid detection of veterinary drug residues, in particular to an immunochromatographic reaction using colloidal gold markers for color development, an immunocolloidal gold test strip for rapid detection of sulfadiazine residues and a preparation method thereof . Background technique [0002] For a long time, sulfadiazine (SD) has been used as an excellent antibacterial drug in clinical and veterinary medicine. Studies in recent years have found that sulfadiazine has many side effects, such as: the crystallization of sulfadiazine and its acetyl derivatives in urine can cause kidney damage, liver damage, terminal neuritis and a disease similar to nodular peripheral arteritis Diseases, as well as drug allergic dermatitis, leukopenia, hemolytic anemia and drug fever. In addition, many bacteria can also develop resistance to it when it is used for a long time. Therefore, at present, sulfadiaz...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/18C07K16/18G01N33/577G01N33/558G01N33/532G01N33/52
Inventor 毕丁仁王喜亮熊宁石德时李自力刘百红李晓云张道宏金秀娥
Owner HUAZHONG AGRI UNIV
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