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Test strip and detection card for detecting PAT/bar protein, preparation method and application of test strip
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A test strip and protein technology, applied in the field of immune analysis, can solve the problems of poor sensitivity, prone to false positives, non-specific interference, etc.
Pending Publication Date: 2021-05-14
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI +1
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Therefore, colloidal gold has at least one of the following disadvantages: ① colloidal gold is a particle visible to the naked eye, and its sensitivity is poor; ② colloidal gold and protein are combined by electrostatic attraction, which has non-specific interference and is prone to false positives ; ③Immune binding colloidal gold chromatography can only be used for qualitative detection, not for quantitative detection of Bar gene
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[0088] In a second aspect, the embodiments of the present invention provide a method for preparing the test strip as described in the first aspect, the preparation method comprising the following steps:
[0089] (1) Prepare the second PAT / bar protein monoclonalantibody labeled with quantum dots and the rabbit IgG labeled with quantum dots, then mix the labeled antibodies and spray the film to obtain a binding pad;
[0090] (2) After diluting the goat anti-rabbit IgG, streak it onto the reaction membrane to form a quality control line, and after diluting the first PAT / bar protein monoclonal antibody, streak it onto the reaction membrane to form a detection line;
[0091] (3) Fixing the sample pad, the binding pad, the reaction film and the absorption pad on the bottom plate in sequence to obtain the test strip.
[0092] Further, in step (1), the spray volume of the spray film is 3-6 μL / cm, for example, the spray volume can be 3 μL / cm, 4 μL / cm, 5 μL / cm or 6 μL / cm, etc., preferabl...
Embodiment 1
[0125] This embodiment provides a test strip for rapid detection of PAT / bar protein. The test strip includes a PVC bottom plate connected to a sample pad, a binding pad, a nitrocellulose membrane (NC membrane) and an absorbent pad.
[0126] In the present embodiment, the composition of the various solutions used is shown in Table 1 below:
[0127] Table 1
[0128]
[0129] The preparation step of described test strip comprises:
[0130] (1) Preparation of sample pads: Soak the sample pads in the sample pad treatment solution for 2 hours, then place them in an oven with humidity <30% and a temperature of 45°C, dry them for 18 hours, and store them sealed at 6°C.
[0131](2) Preparation of the bonding pad: soak the bonding pad in the bonding pad treatment solution for 2 hours, take out the soaked bonding pad, and then place it in an oven with a humidity of <30% and a temperature of 45° C. ℃ sealed storage.
[0142] This embodiment provides a test strip for detecting PAT / bar protein, which is basically the same as the test strip described in Example 1, except that the working concentration of the first PAT / bar protein monoclonal antibody is 0.7mg / mL, the working concentration of the goat anti-rabbit IgG is 0.7mg / mL.
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Abstract
The invention belongs to the technical field of immunoassay, and particularly relates to a test strip and a detection card for detecting PAT / bar protein, and a preparation method and application of the test strip. A detection line and a quality control line are arranged on a reaction film of the test strip, the detection line is coated with a first PAT / bar proteinmonoclonalantibody, and the quality control line is coated with goat anti-rabbit IgG; and a combination pad of the test strip is coated with a second PAT / bar proteinmonoclonalantibody marked by quantum dots and rabbit IgG marked by quantum dots. The test strip provided by the invention is high in sensitivity, strong in accuracy, good in repetition rate, good in specificity and simple and convenient to operate, can realize qualitative or quantitative detection of Bar genes in transgenic plants or foods, provides technical support for rapid qualitative or quantitative detection of herbicide-resistant transgenic ingredients of plants and foods, and is of great significance to detection of transgenic ingredients of agricultural products such as plants and food and safety evaluation of transgenic food.
Description
technical field [0001] The invention belongs to the technical field of immune analysis, and in particular relates to a test strip for detecting PAT / bar protein, a detection card, a preparation method and application thereof. Background technique [0002] Weeds are produced along with human production activities, and their existence is the result of long-term adaptation to climate, soil, crops, farming systems and social factors and competition with cultivated crops. Human beings have been preventing and controlling weeds since the beginning of agricultural production. Spraying herbicides is currently the most efficient way, which liberates human beings from primary labor and greatly improves efficiency. But because herbicides can kill weeds and crops at the same time, the commercialization of genetically modified crops with herbicide resistance genes has become one of the most advantageous traits in agricultural production. As of 2003, herbicide-resistant crops accounted fo...
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