Immune chromatography test paper detection method based on catalyzing and amplifying of detection signal
An immunochromatographic test paper and signal detection technology, which is applied in biological testing, material inspection products, etc., can solve the problems of detection sensitivity, difficulty in distinguishing detection signals, and fuzzy detection signals, so as to achieve easy judgment and preservation, and broad application prospects , fast effect
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Embodiment 1
[0039] Example 1: Determination of human chorionic gonadotropin (HCG) by double-antibody sandwich method
[0040] (1) Preparation of nanomaterials: c(trisodium citrate) / c(H 2 PdCl 4 )=20, c(H 2 PdCl 4 )=0.5mM as an example, add 15ml H to a 100ml round bottom flask 2 PdCl 4 solution (2mM), 0.1784g trisodium citrate, 2ml ethanol and 43mlddH 2 O, with mechanical stirring, heated to reflux at 115°C for 3h. The solution turns from yellow to brown yellow and then turns brown, and the Pd sol stabilized by trisodium citrate is obtained.
[0041] (2) Nanomaterial-labeled antibody: The prepared Pd sol was centrifuged at high speed (13000rpm, 10min), the supernatant was removed, concentrated 5 times and then redissolved in 1ml ddH 2 O, adjust to the optimum pH 9.0, add β-HCG monoclonal antibody, incubate at room temperature for one hour, add 100 μL 10% BSA, react for 30 minutes, centrifuge at 4°C×12000rpm×20min to retain the precipitate, and remove the supernatant. Wash the marke...
Embodiment 2
[0052] Embodiment 2: Determination of benzopyrene by competitive method
[0053] (1) Preparation of nanomaterials: first, 0.0355g PdCl 2 Dissolve 47μL of concentrated hydrochloric acid solution (12M) in 100mL of water, heat and ultrasonically disperse to form 2mM H 2 PdCl 4 solution. Next, weigh 0.045g MUA and 0.1g hydrazine hydrate respectively, place them in a three-necked bottle, and add 15mL of water. After the MUA was completely dissolved, its pH was adjusted to 12 with 0.1M NaOH solution, while the temperature of the solution was controlled at 35°C. Then, 5 mL of the 2 mM H 2 PdCl 4 The solution was slowly dripped into the three-necked bottle (speed 1ml / 5min). As the reaction progressed, the color of the solution gradually changed from colorless to black. After the reaction, the above solution was centrifuged at 13000 rpm for 20 minutes, and after removing the supernatant, it was dispersed in water again. Repeat the above process 5 times to remove excess stabiliz...
Embodiment 3
[0059] Embodiment 3: the preparation of nanometer material
[0060] In the above embodiments, we have only selected two kinds of nanomaterials for specific description. In fact, there are many kinds of nanomaterials that can be applied in the present invention, such as:
[0061] a. Preparation of palladium particles or platinum particles:
[0062] (1) Preparation of palladium particles: c (trisodium citrate) / c (H 2 PdCl 4 )=20, c(H 2 PdCl 4 )=0.5mM as an example, add 15ml H to a 100ml round bottom flask 2 PdCl 4 Solution (2mM), 0.1784g trisodium citrate, 2ml ethanol and 43mlddH2O, mechanically stirred, heated to reflux at 115°C for 3h. The solution turns from yellow to brown yellow and then turns brown, and the Pd sol stabilized by trisodium citrate is obtained.
[0063] (2) Preparation of platinum particles: c (trisodium citrate) / c (H 2 PtCl 4 )=20, c(H 2 PtCl 4 )=0.5mM as an example, add 15ml H to a 100ml round bottom flask 2 PtCl 4 Solution (2mM), 0.1784g triso...
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