阅读说明：本技术 一种四环素类抗生素六联检测卡 (Tetracycline antibiotic six-joint detection card ) 是由 曾令文 陆琼 于 2019-10-16 设计创作，主要内容包括：本发明公开了一种四环素类抗生素六联检测卡,包括平板和位于平板的上表面的胶体金试纸条组,胶体金试纸条组包括第一至第六胶体金试纸条,通过将待测样品溶液滴加至样品垫上进行6种四环素类抗生素的检测。本发明提出的四环素类抗生素六联检测卡能够同时检测6种常见的四环素类抗生素,特异性强,灵敏度高,检出限低,不仅可以应用在动物组织上,也可以对禽蛋类进行检测。(The invention discloses a tetracycline antibiotic six-joint detection card which comprises a flat plate and a colloidal gold test strip group positioned on the upper surface of the flat plate, wherein the colloidal gold test strip group comprises first to sixth colloidal gold test strips, and a sample solution to be detected is dripped on a sample pad to detect 6 tetracycline antibiotics. The tetracycline antibiotic six-joint detection card provided by the invention can be used for simultaneously detecting 6 common tetracycline antibiotics, has strong specificity, high sensitivity and low detection limit, and can be applied to animal tissues and poultry eggs.)

1. The six-joint detection card for the tetracycline antibiotics is characterized by comprising a flat plate and a colloidal gold test strip group, wherein the colloidal gold test strip group is positioned on the upper surface of the flat plate and comprises a first colloidal gold test strip, a second colloidal gold test strip, a third colloidal gold test strip, a fourth colloidal gold test strip, a fifth colloidal gold test strip and a sixth colloidal gold test strip;

the test line of the first colloidal gold test strip is sprayed with a chlortetracycline-BSA full antigen, the control line is sprayed with a goat anti-mouse chlortetracycline secondary antibody, and a colloidal gold-labeled chlortetracycline antibody is fixed on the binding pad; the test line of the second colloidal gold test strip is sprayed with oxytetracycline-BSA full antigen, the control line is sprayed with goat anti-mouse oxytetracycline secondary antibody, and a colloidal gold-labeled oxytetracycline antibody is fixed on the binding pad; tetracycline-BSA full antigen is sprayed at the test line of the third colloidal gold test strip, goat anti-mouse tetracycline secondary antibody is sprayed at the control line, and a colloidal gold-labeled tetracycline antibody is fixed on the binding pad; the test line of the fourth colloidal gold test strip is sprayed with a methacycline-BSA full antigen, the control line is sprayed with a goat anti-mouse methacycline secondary antibody, and a colloidal gold labeled methacycline antibody is fixed on the binding pad; the test line of the fifth colloidal gold test strip is sprayed with doxycycline-BSA full antigen, the control line is sprayed with goat anti-mouse doxycycline secondary antibody, and a colloidal gold labeled doxycycline antibody is fixed on the binding pad; minocycline-BSA full antigen is sprayed at the test line of the sixth colloidal gold test strip, goat anti-mouse minocycline secondary antibody is sprayed at the control line, and a colloidal gold-labeled minocycline antibody is fixed on the binding pad.

2. The detection card of claim 1, wherein the particle size of the colloidal gold in each colloidal gold strip in the set of colloidal gold strips is 10-15 nm.

3. The test card of claim 1, wherein each colloidal gold strip in the set of colloidal gold strips has a width of 2.5-3.5 mm.

4. The test card of claim 1, wherein the length of the absorbent paper, the nitrocellulose membrane and the sample pad in each colloidal gold strip in the colloidal gold strip group is 17-19 mm, and the length of the conjugate pad is 8-10 mm.

5. The test card of claim 4, wherein the length of the absorbent paper, the nitrocellulose membrane and the sample pad in each colloidal gold strip in the set of colloidal gold strips is 18mm, and the length of the conjugate pad is 9 mm.

6. The test card according to claim 5, wherein the total length of the overlapping portions between the absorbent paper, the nitrocellulose membrane, the sample pad and the conjugate pad in each colloidal gold strip in the set of colloidal gold strips is 1.5 to 2 mm.

7. The test card of claim 6, wherein the distance between the control line and the test line in each colloidal gold strip in the set of colloidal gold strips is 9-11 mm.

8. The test card of claim 7, wherein the distance between the control line and the test line in each colloidal gold strip in the set of colloidal gold strips is 10 mm.

Technical Field

The invention relates to the field of antibiotic detection, in particular to a tetracycline antibiotic six-joint detection card.

Background

Tetracyclines (TCs) antibiotics are a broad-spectrum antibiotic produced by actinomycetes, including chlortetracycline, oxytetracycline, tetracycline and semisynthetic derivatives of methacycline, doxycycline and minocycline, and all have tetracene basic skeleton. Tetracycline antibiotics are widely used in the treatment of bacterial infectious diseases because they inhibit the synthesis of bacterial proteins. In actual work, the tetracycline drugs are left in animal-derived tissues due to random increase of the applied dose and non-compliance with the withdrawal period. Tetracycline drugs remain in animal tissues for a long time, indirectly cause drug resistance to be spread among human beings, and seriously damage human health.

Currently, common analytical detection methods for tetracycline drugs in animal food include chromatography, microbiological methods and enzyme-linked immunization. However, these methods have the disadvantages of expensive purchase of the apparatus, complicated sample pretreatment, long analysis time, and no on-site operation, and thus are limited in their applications. In recent years, the colloidal gold chromatography has attracted attention in the field of analysis and detection due to the advantages of simplicity, rapidness, strong specificity, high sensitivity, capability of being judged by naked eyes, no need of special instruments and equipment and the like. The colloidal gold test strip generally comprises a bottom plate, a nitrocellulose membrane, a combination pad, absorbent paper and a sample pad; the nitrocellulose membrane is adhered to the middle part of the bottom plate, one end of the nitrocellulose membrane is adhered with a combination pad, the other end of the nitrocellulose membrane is adhered with absorbent paper, and a sample pad is adhered on the combination pad; a control line and a test line are sequentially arranged on the nitrocellulose membrane from the end close to the water absorption paper; the sample pad is dripped with liquid to be detected and then permeates to the other end, and is combined by the antigen and the antibody, and the aim of detection is fulfilled by utilizing colloidal gold to present color reaction.

At present, few researches on detecting tetracycline antibiotics by using colloidal gold test paper exist, and a colloidal gold chromatography technology capable of simultaneously detecting 6 tetracycline antibiotics does not exist, so that the colloidal gold chromatography technology has significance in simultaneously detecting 6 tetracycline antibiotics.

Disclosure of Invention

The invention provides a tetracycline antibiotic six-joint detection card, which comprises a flat plate and a colloidal gold test strip group, wherein the colloidal gold test strip group is positioned on the upper surface of the flat plate and comprises a first colloidal gold test strip, a second colloidal gold test strip, a third colloidal gold test strip, a fourth colloidal gold test strip, a fifth colloidal gold test strip and a sixth colloidal gold test strip;

the test line of the first colloidal gold test strip is sprayed with a chlortetracycline-BSA full antigen, the control line is sprayed with a goat anti-mouse chlortetracycline secondary antibody, and a colloidal gold-labeled chlortetracycline antibody is fixed on the binding pad; the test line of the second colloidal gold test strip is sprayed with oxytetracycline-BSA full antigen, the control line is sprayed with goat anti-mouse oxytetracycline secondary antibody, and a colloidal gold-labeled oxytetracycline antibody is fixed on the binding pad; tetracycline-BSA full antigen is sprayed at the test line of the third colloidal gold test strip, goat anti-mouse tetracycline secondary antibody is sprayed at the control line, and a colloidal gold-labeled tetracycline antibody is fixed on the binding pad; the test line of the fourth colloidal gold test strip is sprayed with a methacycline-BSA full antigen, the control line is sprayed with a goat anti-mouse methacycline secondary antibody, and a colloidal gold labeled methacycline antibody is fixed on the binding pad; the test line of the fifth colloidal gold test strip is sprayed with doxycycline-BSA full antigen, the control line is sprayed with goat anti-mouse doxycycline secondary antibody, and a colloidal gold labeled doxycycline antibody is fixed on the binding pad; minocycline-BSA full antigen is sprayed at the test line of the sixth colloidal gold test strip, goat anti-mouse minocycline secondary antibody is sprayed at the control line, and a colloidal gold-labeled minocycline antibody is fixed on the binding pad.

The detection is mainly carried out by the principle that the antibiotic and the antigen coated on the detection line compete to combine with the colloidal gold-labeled antibody adsorbed on the binding pad. After the antibiotic positive sample is dripped into the sample pad, the antibiotic positive sample is specifically combined with the antibiotic antibody marked by the colloidal gold on the combination pad to form colloidal gold-antibiotic antibody-antibiotic, and then the antigen attached on the detection line can compete to be combined with the limited antigen sites on the surface of the antibody through the detection line. The more antibiotic in the sample, the more colloidal gold-labeled antibody is bound, resulting in failure of the antigen on the detection line to bind to the colloidal gold-labeled antibiotic antibody. When the colloidal gold labeled antibiotic antibody on the detection line is reduced to a certain degree, the detection line is light or colorless. And (3) continuously carrying out chromatography on the rest of the colloidal gold labeled antibiotic antibody, wherein the goat anti-mouse antibiotic secondary antibody on the control line does not have a competitive site relation with the antibiotic, and when the colloidal gold-antibiotic antibody-antibiotic reaches the control line, the colloidal gold labeled antibiotic antibody-antibiotic is specifically combined with the goat anti-mouse antibiotic secondary antibody coated on the quality control line, so that the control line develops color.

Preferably, the particle size of the colloidal gold in each colloidal gold test strip in the colloidal gold test strip group is 10-15 nm.

Preferably, the width of each colloidal gold test strip in the colloidal gold test strip group is 2.5-3.5 mm.

Preferably, the lengths of the absorbent paper, the nitrocellulose membrane and the sample pad in each colloidal gold test strip in the colloidal gold test strip group are 17-19 mm, and the length of the combination pad is 8-10 mm.

Preferably, the length of the absorbent paper, the length of the nitrocellulose membrane and the length of the sample pad in each colloidal gold test strip in the colloidal gold test strip group are all 18mm, and the length of the combination pad is 9 mm.

Preferably, the total length of the overlapping part among the absorbent paper, the nitrocellulose membrane, the sample pad and the binding pad in each colloidal gold test strip in the colloidal gold test strip group is 1.5-2 mm.

Preferably, the distance between the control line and the test line in each colloidal gold test strip in the colloidal gold test strip group is 9-11 mm.

Preferably, the distance between the control line and the test line in each colloidal gold test strip in the colloidal gold test strip group is 10 mm.

Has the advantages that: the six-joint detection card for the tetracycline antibiotics can be used for simultaneously detecting 6 common tetracycline antibiotics, is high in specificity and sensitivity, the lowest detection limit can reach 5 mu g/kg of aureomycin, 6 mu g/kg of oxytetracycline, 4 mu g/kg of tetracycline, 7 mu g/kg of methacycline, 5 mu g/kg of doxycycline and 6 mu g/kg of minocycline residues, and can be applied to animal tissues and also used for detecting poultry eggs.

Drawings

FIG. 1 is a schematic structural diagram of a colloidal gold test strip in a colloidal gold test strip group according to the present invention;

fig. 2 is a schematic structural diagram of the tetracycline antibiotic detection card of the present invention.

Detailed Description

The concept and technical effects of the present invention will be clearly and completely described in the following embodiments to fully understand the objects, aspects and effects of the present invention. It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict.