CN102109524A - Preparation method of test paper strip for Clenbuterol hydrochloride with high sensitivity - Google Patents

Abstract

The invention relates to a preparation method of a test paper strip for Clenbuterol hydrochloride with high sensitivity. The preparation method comprises the following steps: uniformly coating colloidal gold-Clenbuterol hydrochloride monoclonal antibodies on glass fibers, uniformly spraying Clenbuterol hydrochloride antigens and goat-anti-mouse IgG antibodies on a nitrocellulose membrane as a detecting line and a quality control line, then arranging the glass fibers, gold-marked Clenbuterol hydrochloride monoclonal antibody glass fibers, the nitrocellulose membrane and hard water-absorbing filter paper from bottom to top, adhering the glass fibers, the gold-marked Clenbuterol hydrochloride monoclonal antibody glass fibers, the nitrocellulose membrane and the hard water-absorbing filter paper on a single-faced glue PVC (polyvinyl chloride) board side, and cutting the PVC board into paper strips longitudinally. Compared with the prior art, in the test paper strip prepared by the invention for detection, the detection sensitivity is high, the minimum detection amount is 1ppb, and the preparation method is simple and convenient.

Description

A kind of preparation method of high-sensitivity clenbuterol hydrochloride detection test strip

technical field

The invention relates to a preparation method of a detection test strip, in particular to a preparation method of a high-sensitivity clenbuterol hydrochloride detection test strip.

Background technique

Clenbuterol (CL), also known as dichlorohydrin, is a β ₂ receptor agonist, which can effectively improve animal meat production performance and feed reward. However, it has toxic side effects on internal organs such as liver and kidney of humans and animals, and can cause death in serious cases. The Ministry of Agriculture clearly stipulates that clenbuterol and other β ₂ receptor agonists are prohibited from being used as feed additives. Enzyme-linked immunosorbent assay (ELISA) is a common method for quantitative detection of CL at present, suitable for laboratory operation, but not convenient for on-site detection (such as farms, feed processing plants, slaughterhouses, pork sales markets) . Colloidal gold test strips are more suitable for on-site qualitative testing. The detection sensitivity of colloidal gold test strips currently on the market is poor, and on-site missed detection often occurs.

Contents of the invention

The object of the present invention is to provide a kind of preparation method of the high-sensitivity clenbuterol hydrochloride detection test strip with high detection sensitivity and easy to use in order to overcome the defects in the above-mentioned prior art.

The purpose of the present invention can be achieved through the following technical solutions:

A kind of preparation method of high-sensitivity clenbuterol hydrochloride detection test strip is characterized in that, the method comprises the following steps:

(1) Apply the colloidal gold-clenbuterol monoclonal antibody evenly on the glass fiber with a micropipette, and then vacuum-dry it at 20-25°C;

(2) Evenly spray clenbuterol hydrochloride antigen and goat anti-mouse IgG antibody on the nitrocellulose membrane as the detection line and quality control line, and dry the nitrocellulose membrane at 37°C for 2-3 hours;

(3) the nitrocellulose membrane is cut into strips with a width of 5-6mm, and placed in a dry environment for subsequent use;

(4) Arrange and adhere glass fiber, gold-labeled clenbuterol hydrochloride glass fiber, solidified clenbuterol hydrochloride antigen and goat anti-mouse IgG antibody nitrocellulose membrane and hard absorbent filter paper from bottom to top On one side of the single-sided adhesive PVC board, then cut the assembled PVC board lengthwise into test strips.

The size of the nitrocellulose membrane in the step (2) is 80mm×300mm.

In the step (2), the detection line is set below the quality control line.

The detection test strip size in the step (4) is 5mm×80mm.

Compared with the prior art, the detection test strip prepared by the invention has high detection sensitivity, the minimum detection amount is 1ppb, and the preparation method is simple and convenient.

Description of drawings

Fig. 1 is the assembly schematic diagram among the embodiment 1;

Fig. 2 is the schematic diagram that detects test strip in embodiment 1.

In the figure, 1 is glass fiber, 2 is gold-labeled clenbuterol hydrochloride glass fiber, 3 is nitrocellulose membrane, 31 is detection line, 32 is quality control line, 4 is hard absorbent filter paper, 5 is single-sided adhesive PVC board.

Detailed ways

The present invention will be described in detail below in conjunction with the accompanying drawings and specific embodiments.

Example 1

A kind of preparation method of high-sensitivity clenbuterol hydrochloride detection test strip, the method comprises the following steps:

(1) Colloidal gold-clenbuterol hydrochloride monoclonal antibody is evenly coated on the glass fiber 2 with a size of 80 mm × 300 mm with a micropipette tip, and then vacuum-dried at room temperature of 20 ° C. The glass fiber was purchased from Shanghai Gold Standard Biotechnology Co., Ltd., for SB06 glass fiber;

(2) Spray the clenbuterol hydrochloride antigen and goat anti-mouse IgG antibody evenly on the nitrocellulose membrane 3 with a size of 80mm×300mm as the detection line 31 and the quality control line 32, and the detection line 31 is set below the quality control line 32 , and then dry the nitrocellulose membrane 3 at 37°C for 2h;

(3) Cut the nitrocellulose membrane 3 into strips with a width of 5-6mm with a paper cutter, and place it in a dry environment for subsequent use;

(4) Glass fiber 1, gold-labeled clenbuterol hydrochloride glass fiber 2, nitrocellulose membrane 3 of solidified clenbuterol hydrochloride antigen and goat anti-mouse IgG antibody, and rigid water-absorbing filter paper 4 are bottom-up Arrange it as shown in Figure 1, and then stick it to one side of the single-sided adhesive PVC board 5, and then use a strip cutter to cut the assembled PVC board longitudinally into 5mm×80mm test strips. as shown in picture 2.

The quality control line 32 of the assembled test strip turns red during the test, indicating that the test strip is qualified. If the test line 31 changes color during the test, it indicates that there is clenbuterol hydrochloride (CL) in the sample to be tested. Line 31 does not change color, indicating that the sample to be tested contains CL.

Example 2

A kind of preparation method of high-sensitivity clenbuterol hydrochloride detection test strip, the method comprises the following steps:

(1) Colloidal gold-CL monoclonal antibody is evenly coated on the glass fiber with a micropipette tip, and then vacuum-dried at room temperature of 25°C;

(2) Spray clenbuterol hydrochloride antigen and goat anti-mouse IgG antibody evenly on the nitrocellulose membrane with a size of 80mm×300mm as the detection line and quality control line. The detection line is set below the quality control line, and then the nitrocellulose The film was dried at 37°C for 3 hours;

(3) the nitrocellulose membrane is cut into strips of 5-6 mm, and placed in a dry environment for subsequent use;

(4) Arrange glass fiber, gold-labeled clenbuterol hydrochloride glass fiber, solidified clenbuterol hydrochloride antigen and goat anti-mouse IgG antibody nitrocellulose membrane and hard absorbent filter paper from bottom to top, Then stick it to one side of the single-sided adhesive PVC board, and then cut the assembled PVC board longitudinally into 5mm×80mm test strips.

The quality control line of the assembled test strip turns red during the test, indicating that the quality of the test strip is qualified. If the test line changes color during the test, it indicates that there is clenbuterol hydrochloride (CL) in the sample to be tested. If the test line is not Discoloration indicates that CL is contained in the sample to be tested.

Claims (4)

1. a kind of preparation method of high-sensitivity clenbuterol hydrochloride detection test strip is characterized in that, the method may further comprise the steps: (1) Colloidal gold-clenbuterol hydrochloride monoclonal antibody is evenly coated on the glass fiber with a micropipette tip, and then vacuum-dried at 20-25°C; (2) Evenly spray clenbuterol hydrochloride antigen and goat anti-mouse IgG antibody on the nitrocellulose membrane as the detection line and quality control line, and dry the nitrocellulose membrane at 37°C for 2-3 hours; (3) the nitrocellulose membrane is cut into strips with a width of 5-6mm, and placed in a dry environment for subsequent use; (4) Arrange and adhere glass fiber, gold-labeled clenbuterol hydrochloride glass fiber, solidified clenbuterol hydrochloride antigen and goat anti-mouse IgG antibody nitrocellulose membrane and hard absorbent filter paper from bottom to top On one side of the single-sided adhesive PVC board, then cut the assembled PVC board lengthwise into test strips. 2. the preparation method of a kind of high-sensitivity clenbuterol hydrochloride detection test strip according to claim 1, is characterized in that, the nitrocellulose membrane size in the described step (2) is 80mm * 300mm. 3. the preparation method of a kind of high-sensitivity clenbuterol hydrochloride detection test paper strip according to claim 1, is characterized in that, in described step (2), detection line is located at the below of quality control line. 4. the preparation method of a kind of high-sensitivity clenbuterol hydrochloride detection test strip according to claim 1, is characterized in that, the detection test strip size in the described step (4) is 5mm * 80mm.

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