Test paper card for synchronously detecting zearalenone and aflatoxin B1, and preparation and detection method

A technology of zearalenone and aflatoxin, which is applied in the field of immunological detection, can solve the problems that cannot be applied to actual production and life, long detection cycle, cumbersome operation, etc., and achieve the effects of high sensitivity, shortened time, and improved accuracy

Inactive Publication Date: 2018-05-01
洛阳现代生物技术研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection methods of aflatoxin B1 and zearalenone include thin film chromatography and liquid chromatography. Although these anal

Method used

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  • Test paper card for synchronously detecting zearalenone and aflatoxin B1, and preparation and detection method
  • Test paper card for synchronously detecting zearalenone and aflatoxin B1, and preparation and detection method
  • Test paper card for synchronously detecting zearalenone and aflatoxin B1, and preparation and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1, test paper card preparation

[0025] 1. Preparation of fluorescent microsphere-antibody complexes

[0026] (1) Cleaning: Take 50 μL of fluorescent microspheres into a 1.5 mL centrifuge tube, add 1 mL of 0.01M MES buffer, shake and mix, centrifuge at 15,000 r / min for 15 min, discard the supernatant, add 1 mL of 0.01M MES buffer, and ultrasonically Loose microspheres; repeat this step three times, the purpose of cleaning the microspheres has been achieved, and finally the microsphere liquid after cleaning is obtained;

[0027] (2) Activation: Add 250 μL of EDC solution to 1 mL of microsphere solution after washing, activate in the dark for 2 h, centrifuge at 15,000 r / min for 15 min, discard the supernatant, add 1 mL of 0.01M MES buffer, and disperse the microspheres by ultrasonication, 15,000 r / min Centrifuge for 15min, discard the supernatant;

[0028](3) Labeling: add 1mL 0.05M borate buffer, ultrasonically break up, add 8μg zearalenone monoclonal antib...

Embodiment 2

[0045] Embodiment 2, drawing of standard curve

[0046] 1. Configuration of standard products:

[0047] Preparation of zearalenone gradient standard: Dilute zearalenone standard to 0.2ppb, 0.4ppb, 0.8ppb, 1.6ppb, 3.2ppb with 0.1M pH7.4 phosphate buffer;

[0048] Preparation of aflatoxin B1 gradient standard: Dilute aflatoxin B1 standard to 0.2ppb, 0.4ppb, 0.8ppb, 1.6ppb, 3.2ppb with 0.1M pH7.4 phosphate buffer;

[0049] 2. Reading:

[0050] Point the above-mentioned standard product on the card, react for 10 minutes, and read it in the corresponding card slot of the portable fluorescence immunoassay analyzer. The reading results are shown in Table 1 below:

[0051]

[0052] 3. Verification of the calculation method:

[0053] (1) The four-parameter method, after inputting the corresponding parameters of the zearalenone curve into the portable fluorescence immunoassay analyzer, randomly select two points of 0.4ppb and 1.6ppb in the gradient standard, click the card, react ...

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Abstract

The invention relates to a test paper card for synchronously detecting zearalenone and aflatoxin B1, and a preparation and a detection method, and belongs to the field of immunology detection. The test paper card comprises a card shell and a test paper strip, wherein the test paper strip comprises a bottom plate, a water absorption pad, a detection pad, a combination pad and a sample pad; the water adsorption pad, the detection pad, the combination pad and the sample pad are overlapped and pasted to the bottom plate in sequence; the detection pad is an NC membrane (nitrocellulose filter membrane) provided with a quality control line C, a detection line T1 and a detection line T2; the quality control line C coats a goat-anti-mouse second antibody, the detection line T1 coats ZEN-BSA, and the detection line T2 coats AFB1-BSA; the combination pad is a glass cellulose membrane which embeds an anti-zearalenone monoclonal antibody marked by time resolution fluorescent microspheres and anti-aflatoxin B1 monoclonal antibody; the sample pad is the glass cellulose membrane obtained in a way that drying is carried out after the dipping processing of sample pad conditioning fluid. The invention provides a new method for systematically, conveniently, normatively, quantitatively and synchronously detecting the zearalenone and the aflatoxin, stability is good, and sensitiveness is high.

Description

technical field [0001] The invention belongs to the field of immunological detection, and in particular relates to a test paper card for synchronous detection of zearalenone and aflatoxin, a preparation method and a detection method. Background technique [0002] Zearalenone and aflatoxin B1 are two common mycotoxins, and the concentration of these two toxins in feed is closely related to the prevalence of animal epidemics. Zearalenone (F-2) is a secondary metabolite produced by Gibberella. It was originally isolated from corn with Gibberella. It has estrogenic effects and can cause acute and chronic poisoning in animals, thereby causing abnormal reproductive functions in animals. Cause death and cause huge economic losses to livestock farms. Aflatoxin B1 (AFB1), as the most carcinogenic chemical substance known, is 30 times stronger than vomitoxin (Don) and 25 times stronger than F-2; Reduced weight gain and feed conversion, pulmonary edema, infertility or abortion, incre...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 原小燕刘近张军玲赵津子杜小波刘明瑞
Owner 洛阳现代生物技术研究院有限公司
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