Colloidal gold chromatography test paper strip for quickly detecting bisphenol A and preparation method thereof

Abstract

The invention discloses a colloidal gold chromatography test paper strip for quickly detecting bisphenol A and a preparation method thereof, and belongs to the technical field of detection of biological immunization methods. The detecting test paper strip consists of a lining board, and a sample pad, a gold labeled combining pad, an enveloping film and a water absorbing pad, which are orderly connected on the lining board, wherein the gold labeled combining pad is glass fiber wool adsorbing gold-labeled antibodies of the bisphenol A; and the enveloping film is provided with a linear invisible test line T line which is printed by carrier protein solution coupled by diphenolic acid, and a linear invisible contrast line C line which is printed by sheep anti-rabbit IgG solution, and the two lines are parallel. The test paper strip has strong specificity, high sensibility with the lowest detection limit of 5ppb, simplicity, convenience, quickness and strong timeliness, does not need any other reagents and instruments, can be operated in field, can determine a detection result in 15 minutes after being put into tested sample solution, has vivid, directviewing and accurate result display, saves cost, has wide applicable range and contributes to promotion.

Description

Technical field

[0001] A colloidal gold chromatography test strip for rapid detection of bisphenol A and a preparation method thereof belong to the technical field of detection of biological immune methods. Background technique

[0002] Bisphenol A (BPA) is a kind of "Endocrine Disrupting Chemicals (ECD)" newly discovered from food packaging materials in recent years and has certain estrogenic properties. The discovery of BPA has aroused widespread interest in food and packaging materials in various countries. There have been reports of BPA being detected from canned vegetables, baby preparations, and food cans. BPA is the main raw material for the production of PC resin and epoxy (EP) resin. It is also used in phenolic resin, plastic polyester, antioxidant and polyvinyl chloride (PVC) stabilizer. Therefore, it has important applications in food packaging materials and container inner wall coatings. In occupational production and daily life, BPA can enter the human body through t...

Examples

preparation example Construction

[0036] 2. Preparation of anti-BHPVA polyclonal antibody

[0037] Preparation of polyclonal antibodies: New Zealand white rabbits were immunized with BHPVA-BSA antigen at a dose of 1 mg / time, and injected subcutaneously at multiple points on the back. For the first immunization, mix BHPVA-BSA antigen with the same amount of Freund’s complete adjuvant (FCA) to fully emulsify; boost immunization, use BHPVA-BSA antigen to mix with the same amount of Freund’s incomplete adjuvant (FIA), fully emulsify, Three weeks after the immunization, 4-5 consecutive immunizations, with an interval of 2 to 3 weeks, and 10 to 15 days after the last immunization, the titer reached 10 by ELISA. 5 At the above, blood was collected and serum was separated and collected. The IgG antibody was extracted by saturated ammonium sulfate salting out method, and stored at -20°C for later use.

[0038] 3. Preparation of BHPVA gold-labeled antibody and gold-labeled substance binding pad

[0039] Preparation of collo...

Embodiment 1

[0042] Example 1: Spray 0.5 mg / mL of coating antigen and goat anti-rabbit IgG on the coating film as the test line (T) and control line (C), and dry in an oven at 37°C for 10 minutes. In the same way, a certain concentration of gold-labeled antibody is coated on the binding pad. The test strip is composed of a lining board, on which the sample pad, colloidal gold bonding pad, coating film and absorbent pad are glued in order. Cut the pasted board into 3mm wide strips, and then put the test strips and desiccant into an aluminum foil bag and seal for storage.

[0043] Detection operation method: insert the sample end of the bisphenol A colloidal gold chromatography test strip into the sample solution to be tested, the insertion depth does not exceed the mark line, take out the test strip for about 10-20 seconds, and place it horizontally for about 15 minutes Observe to determine the test results.

[0044] Result judgment: If only one red line of C line appears on the coating film, i...

Technical field

[0001] A colloidal gold chromatography test strip for rapid detection of bisphenol A and a preparation method thereof belong to the technical field of detection of biological immune methods. Background technique

[0002] Bisphenol A (BPA) is a kind of "Endocrine Disrupting Chemicals (ECD)" newly discovered from food packaging materials in recent years and has certain estrogenic properties. The discovery of BPA has aroused widespread interest in food and packaging materials in various countries. There have been reports of BPA being detected from canned vegetables, baby preparations, and food cans. BPA is the main raw material for the production of PC resin and epoxy (EP) resin. It is also used in phenolic resin, plastic polyester, antioxidant and polyvinyl chloride (PVC) stabilizer. Therefore, it has important applications in food packaging materials and container inner wall coatings. In occupational production and daily life, BPA can enter the human body through t...

Examples

preparation example Construction

[0036] 2. Preparation of anti-BHPVA polyclonal antibody

[0037] Preparation of polyclonal antibodies: New Zealand white rabbits were immunized with BHPVA-BSA antigen at a dose of 1 mg / time, and injected subcutaneously at multiple points on the back. For the first immunization, mix BHPVA-BSA antigen with the same amount of Freund’s complete adjuvant (FCA) to fully emulsify; boost immunization, use BHPVA-BSA antigen to mix with the same amount of Freund’s incomplete adjuvant (FIA), fully emulsify, Three weeks after the immunization, 4-5 consecutive immunizations, with an interval of 2 to 3 weeks, and 10 to 15 days after the last immunization, the titer reached 10 by ELISA. 5 At the above, blood was collected and serum was separated and collected. The IgG antibody was extracted by saturated ammonium sulfate salting out method, and stored at -20°C for later use.

[0038] 3. Preparation of BHPVA gold-labeled antibody and gold-labeled substance binding pad

[0039] Preparation of collo...

Embodiment 1

[0042] Example 1: Spray 0.5 mg / mL of coating antigen and goat anti-rabbit IgG on the coating film as the test line (T) and control line (C), and dry in an oven at 37°C for 10 minutes. In the same way, a certain concentration of gold-labeled antibody is coated on the binding pad. The test strip is composed of a lining board, on which the sample pad, colloidal gold bonding pad, coating film and absorbent pad are glued in order. Cut the pasted board into 3mm wide strips, and then put the test strips and desiccant into an aluminum foil bag and seal for storage.

[0043] Detection operation method: insert the sample end of the bisphenol A colloidal gold chromatography test strip into the sample solution to be tested, the insertion depth does not exceed the mark line, take out the test strip for about 10-20 seconds, and place it horizontally for about 15 minutes Observe to determine the test results.

[0044] Result judgment: If only one red line of C line appears on the coating film, i...

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