Novel coronavirus detection kit and preparation method thereof
A detection kit and coronavirus technology, applied in the field of medical testing, can solve the problems of low sensitivity of immune detection, and achieve the effects of strong specificity, low detection cost and high sensitivity
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Embodiment 1
[0044] A detection kit for novel coronavirus
[0045] Among them, the structure of the immunochromatographic surface-enhanced Raman spectroscopy test strip for detecting the new coronavirus is as follows: figure 1 and figure 2 As shown, it includes sample pad 2, binding pad 3, nitrocellulose membrane 4, absorbent pad 5 and PVC backing 1.
[0046] A sample pad, a binding pad, a nitrocellulose membrane and a water-absorbing pad are successively adhered to the PVC backing, the binding pad and the nitrocellulose membrane are overlapped, and the nitrocellulose membrane and the water-absorbing pad are overlapped; The binding pad was coated with Wu-DTNB@Ag-Ab1 marker, and the nitrocellulose membrane was coated with a detection line T composed of the new crown N protein antibody 2 and a quality control line C composed of rabbit anti-mouse IgG, respectively.
[0047] The coating concentration of the new crown N protein antibody 2 is 0.5 mg / mL; the coating concentration of the rabbi...
Embodiment 2
[0051] A kind of preparation method of novel coronavirus detection kit
[0052] (1) Preparation of nano-silver particle solution: Weigh 0.018g of silver nitrate and dissolve it in 100mL of ultrapure water, place the prepared silver nitrate solution in an oil bath (130°C) and heat while stirring. When the solution is boiled, add 4 mL of 1% trisodium citrate solution, continue to heat for 1 hour, and then cool to 20-25° C. to obtain nano-silver particle solution (AgNPs);
[0053] (2) Surface modification of DTNB on silver nanoparticles: take 10 mL of the silver nanoparticles solution prepared in step (1), add 20 μL of 0.01 mol / L DTNB solution, stir and react at room temperature for 2 h, 12000 r / min, centrifuge for 10 min, and remove the supernatant solution, and the precipitation was suspended with 2 mL of ultrapure water to obtain the AgNPs-DTNB solution;
[0054] (3) Preparation of tungsten-wrapped AgNPs-DTNB core-shell structure: take 2 mL of the AgNPs-DTNB solution prepared...
Embodiment 3
[0061] Establishment of standard curve
[0062] Take the new crown recombinant N protein and dilute it with the sample extract to obtain concentrations of 1pg / mL, 10pg / mL, 100pg / mL, and 500pg / mL. After testing with the above test strips, it is detected by confocal laser Raman spectrometer. The characteristic peak intensity of the T-line Raman signal molecule DTNB is used to draw the standard curve, see image 3 .
[0063] technical effect
[0064] specific test
[0065] The pathogenic bacteria or recombinant proteins used for specific tests include SARS-N protein, MERS-N protein, and H1N1 virus isolates.
[0066] Dilute the concentration of the analyte to 100ng / mL, accurately draw 80 μL of the sample to be tested with a pipette, and drop it vertically and slowly into the sample hole of the test card. Start timing after adding the sample, and read the result 10 minutes after adding the sample. The test results are shown in Table 1:
[0067] Table 1 test strip specificity t...
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