Preparation method of coronavirus rapid detection kit and rapid detection equipment

Abstract

The invention provides a preparation method of a rapid coronavirus detection kit and rapid detection equipment, and solves the technical problems that the existing coronavirus detection time is long, and rapid detection of coronavirus cannot be realized, the preparation method of the rapid coronavirus detection kit comprises the following steps: (1) preparing a reagent A, the preparation method comprises the following steps: mixing bovine serum albumin, a buffer solution I, a coagulant and a preservative, and adjusting the pH value to obtain a reagent A with the pH value of 8.0; (2) preparing a reagent B, washing polystyrene microspheres with a buffer solution II, carrying out ultrasonic resuspension, adding EDAC and NHS for activation, and centrifuging to obtain a precipitate II; carrying out ultrasonic resuspension in the buffer solution III, adding a coronavirus antibody, and centrifuging to obtain a precipitate III; carrying out ultrasonic resuspension in the mixed solution III, and centrifuging to obtain a precipitate IV; adding trehalose and glycerol into the buffer solution V to obtain a mixed solution IV; adding the precipitate IV into the mixed solution IV, and carrying out ultrasonic resuspension to obtain a reagent B; the invention further provides equipment for rapidly detecting the coronavirus. The method can be widely applied to the technical field of virus detection.

Description

Technical Field

[0001] This application relates to the technical field of virus detection, and particularly relates to a preparation method of a rapid detection kit for coronavirus and a rapid detection device. Background Art

[0002] In the global public health field, the spread of viruses has always been a severe challenge. Taking coronavirus as a typical representative, it has had a huge impact on human health and social life. At present, nucleic acid detection is the main diagnostic method for coronavirus. It is widely used in China due to its high sensitivity and good specificity. For example, the invention with the publication number CN117126964A discloses a novel coronavirus nucleic acid detection kit, and the entire detection reaction process can be completed within 40 minutes, with simple, fast and highly specific detection.

[0003] However, due to the high requirements for laboratory environment, detection personnel, instruments, etc. in nucleic acid detection, and the long detection time, it cannot achieve rapid and timely detection of viruses, which greatly limits the on-site automatic detection of viruses in public places such as airports and stations. Once faced with a situation where the number of infected people is large and increasing rapidly, it is difficult to meet the diagnostic needs of a large number of suspected infected people and quarantined observers, resulting in a delay in the quarantine time, delaying the travel of passengers, and even posing a risk of a wider range of infections. Therefore, it is imperative to research a new rapid detection kit and rapid detection device for coronavirus. Summary of the Invention

[0004] The purpose of the present invention is to solve the above-mentioned technical deficiencies, and provide a preparation method of a rapid detection kit for coronavirus and a rapid detection device, so as to shorten the detection time of coronavirus and achieve rapid detection of coronavirus.

[0005] To this end, the present invention provides a preparation method of a rapid detection kit for coronavirus, including the following steps:

[0006] S1. Prepare Reagent A

[0007] Mix bovine serum albumin (BSA), Buffer I, coagulant promoter, and preservative, and adjust the pH to finally obtain Reagent A with a pH of 8.0; wherein, the prepared Reagent A contains 2% (by mass) of bovine serum albumin (BSA), 2.4 g / L of Buffer I, 1 - 5 g / L of coagulant promoter, and 0.5 - 5 g / L of preservative;

[0008] S2. Prepare Reagent B

[0009] ① Adjust the pH of Buffer II to obtain a Buffer II solution with a pH of 6.0; Take a part of the Buffer II solution, add polystyrene microspheres with a diameter of 80 - 300 nm for washing. After washing, centrifuge and discard the supernatant, and retain the precipitate I containing polystyrene microspheres for standby;

[0010] ② Take another part of the buffer solution II prepared in step ①, add the precipitate I obtained in step ① thereto, and resuspend it by ultrasonic treatment to make the concentration of polystyrene microspheres in the buffer solution II the preset concentration; then add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDAC) and N-hydroxysuccinimide (NHS) thereto to form a mixed solution I, wherein the concentration of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 2 g / L and the concentration of N-hydroxysuccinimide is 3 g / L; activate the mixed solution I at room temperature, centrifuge to remove the supernatant, and obtain a precipitate II containing polystyrene microspheres for standby;

[0011] ③ Adjust the pH of buffer solution III to obtain a buffer solution III with a pH of 7.0; ultrasonically resuspend the precipitate II obtained in step ② in the buffer solution III to form a mixed solution II; add a coronavirus antibody to the mixed solution II to make the concentration of the coronavirus antibody in the mixed solution II 0.5 - 3 mg / ml, stir and shake well, centrifuge to remove the supernatant, and retain a precipitate III containing polystyrene microspheres for standby;

[0012] ④ Adjust the pH of buffer solution IV, and then add bovine serum albumin (BSA) thereto to obtain a mixed solution III with a pH of 7.4; ultrasonically resuspend the precipitate III obtained in step ③ in the mixed solution III, stir and shake well, centrifuge to remove the supernatant, and retain a precipitate IV containing polystyrene microspheres for standby;

[0013] ⑤ Add trehalose and glycerol to buffer solution V to obtain a mixed solution IV; add the precipitate IV obtained in step ④ to the mixed solution IV and ultrasonically resuspend it to finally obtain reagent B.

[0014] Preferably, in step (1) and step (2), the corresponding pH is adjusted by using NaOH solution and HCl solution.

[0015] Preferably, buffer solution I, buffer solution II, buffer solution III, buffer solution IV, and buffer solution V are respectively one of tris(hydroxymethyl)aminomethane-hydrochloric acid (Tris) buffer solution, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer solution, phosphate buffer solution, acetate buffer solution, glycine buffer solution, and 2-(N-morpholino)ethanesulfonic acid (MES) buffer solution.

[0016] Preferably, the coagulant includes one of PEG 6000 and PEG 8000.

[0017] Preferably, the preservative includes one of sodium azide, thimerosal sodium, phenol, ethylparaben, and hydroxybenzoic acid.

[0018] Preferably, in step ①, in the buffer solution II, the concentration of buffer solution II is 50 mM.

[0019] Preferably, in step ②, the concentration of polystyrene microspheres in the buffer solution II is 2.5 g / L.

[0020] Preferably, in step ②, the precipitate I is ultrasonically resuspended in the buffer solution II, and the ultrasonic power is controlled at 60%; the mixed solution I is activated at room temperature for 20 - 40 min.

[0021] Preferably, in step ③, in the buffer solution III, the concentration of buffer solution III is 30 mM.

[0022] Preferably, in step ③, the precipitate II is ultrasonically resuspended in the buffer solution III at a temperature of 20°C, and the ultrasonic power is controlled at 60%; the stirring speed is 100 - 300 r / min, oscillated for 1 - 4 hours, and the centrifugation speed is 15000 r / min.

[0023] Preferably, in step ④, in the mixed solution III, the concentration of buffer solution IV is 50 mM, and the concentration of bovine serum albumin is 20 g / L.

[0024] Preferably, in step ④, the precipitate III is ultrasonically resuspended in the mixed solution III, where the temperature is controlled at 25°C, the ultrasonic power is controlled at 30% - 60%, the stirring speed is controlled at 120 - 300 r / min, oscillated and sealed overnight, and the centrifugation speed is 15000 r / min.

[0025] Preferably, in step ⑤, in the mixed solution IV, the concentration of buffer solution V is 30 mM, the concentration of trehalose is 35 g / L, and the concentration of glycerol is 35 g / L.

[0026] Preferably, in step ⑤, the precipitate IV is ultrasonically resuspended in the mixed solution IV, and the ultrasonic power is controlled at 30% - 60%.

[0027] A rapid detection device for coronavirus. The on-site rapid detection device for virus is provided with a light source, and also provided with a slit, a grating, a detection mechanism, and a spectrometer. The detection mechanism is provided with a sample bottle for containing the liquid reagent to be detected; the light source outputs a stable beam with a fixed wavelength. The stable beam enters the grating after passing through the slit, is split by the grating and then passes through the sample bottle and the liquid reagent to be detected inside to form a transmitted beam, and the transmitted beam enters the spectrometer for detecting the light intensity; the width of the slit is adjustable, and the adjustment range is 0 - 5 mm; the detection mechanism is also provided with a platform base for placing the sample bottle.

[0028] The invention has the beneficial effects as follows: the invention provides a method for preparing a coronavirus rapid detection kit, wherein reagent A is prepared, bovine serum albumin, buffer I, a coagulant, and a preservative are mixed, and the pH is adjusted to obtain reagent A with a pH of 8.0; reagent A contains 2% BSA, 2.4 g / L buffer I, 1-5 g / L coagulant, and 0.5-5 g / L preservative; reagent B is prepared, polystyrene microspheres are washed and ultrasonically resuspended in buffer II, activated with EDAC and NHS, and centrifuged to obtain precipitate II; ultrasonically resuspended in buffer III solution, coronavirus antibodies are added, and precipitate III is centrifuged to obtain precipitate III; ultrasonically resuspended in mixed solution III, and precipitated to obtain precipitate IV; trehalose and glycerol are added to buffer V to obtain mixed solution IV; precipitate IV is added to mixed solution IV, and ultrasonically resuspended to obtain reagent B. When used, after the sample to be tested and reagent A are mixed, reagent B is added, and detection is performed after incubation. The operation is simple, the detection time is short, and the detection is completed within 5 minutes, so as to improve the efficiency of coronavirus screening.

[0029] The present invention also provides a coronavirus rapid detection device, which is provided with a light source, and a stable light beam of a fixed wavelength generated by the light source obtains a light beam of suitable intensity after passing through a slit, and then obtains a high-purity monochromatic light beam after passing through a sample bottle and the liquid reagent to be tested inside it. The monochromatic light beam generates a transmitted light beam after passing through the sample bottle and the liquid reagent to be tested inside it, and the transmitted light beam enters the spectrometer and can be converted into an intensity signal to obtain the intensity value of the transmitted light beam detected by the spectrometer. The turbidity of the liquid reagent to be tested in the sample bottle is positively correlated with the number of viruses, and the turbidity of the liquid reagent to be tested and the intensity value of the transmitted light beam detected by the spectrometer are correlated, that is, as the turbidity of the liquid reagent to be tested increases, the intensity of the transmitted light beam formed by the light beam of the same intensity passing through the sample bottle and the liquid reagent to be tested inside it is lower; the intensity value of the transmitted light beam is detected by the spectrometer, and the intensity value of the light beam detected is checked against the corresponding value of the virus number-light intensity value standard curve, so that the virus number can be obtained, and the sample detection is completed. The present invention does not rely on existing professional testing sites and can be installed in public places such as airports and stations for on-site rapid virus detection. For example, when detecting coronavirus, the test can be carried out on-site, greatly shortening the detection process, thereby meeting the diagnosis needs of a large number of suspected infected persons and quarantined observers. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the embodiments or the description of the prior art are briefly introduced below.

[0031] Figure 1 This is the light intensity test diagram of the sample to be tested in Example 1;

[0032] Figure 2 This is the light intensity test diagram of the sample to be tested in Example 2;

[0033] Figure 3 Light intensity test diagram of the sample to be tested in Example 3;

[0034] Figure 4 Light intensity test diagram of the sample to be tested in Example 4;

[0035] Figure 5 Light intensity test diagram of the sample to be tested in Example 5;

[0036] Figure 6 Virus quantity - light intensity value standard curve in Example 2;

[0037] Figure 7 Structural schematic diagram of the front view of the present invention;

[0038] Figure 8 For Figure 7 Structural schematic diagram of the perspective view shown;

[0039] Figure 9 For Figure 8 Structural schematic diagram of the perspective view with part of the outer shell removed from another perspective shown;

[0040] Figure 10 For Figure 9 Structural schematic diagram of the enlarged view of part A shown;

[0041] Figure 11 Structural schematic diagram of the sample bottle;

[0042] Figure 12 Structural schematic diagram of the platform base;

[0043] Figure 13 For Figure 12 Structural schematic diagram of the A - A cross - sectional view shown;

[0044] Figure 14 For Figure 12 Structural schematic diagram of the perspective view shown;

[0045] Figure 15 Structural schematic diagram of another form of the platform base;

[0046] Figure 16 Structural schematic diagram of the front view of the detection cover;

[0047] Figure 17 For Figure 16 Structural schematic diagram of the bottom view shown;

[0048] Figure 18 For Figure 17 Structural schematic diagram of the B - B cross - sectional view shown;

[0049] Figure 19 Working principle diagram of the intelligent control system of the present invention.

[0050] Markings in the figure: 1. Light source, 2. Slit, 3. Grating, 4. Spectrometer, 5. Sample bottle, 6. Platform base, 7. Stepper motor, 8. Ring rack, 9. Ring bracket, 10. Support arm, 11. Gear, 12. Slide block, 13. Circumferential slideway, 14. Heat sink, 15. Semiconductor refrigeration chip, 16. Orifice plate, 17. Support part, 18. Blocking and pushing part, 19. Leakage hole, 20. C-shaped notch, 21. V-shaped opening, 22. Infrared sensor, 23. Temperature sensor, 24. Intelligent control device, 25. First mounting blind hole, 26. Second mounting blind hole, 27. Elastic sheet, 28. Bottle body, 29. Bottle cap, 30. Rounded corner structure, 31. Waste sample box, 32. Detection cover, 33. Sample bottle channel, 34. Platform base channel, 35. Light beam through hole, 36. Outer shell, 37. Bottle placing window, 38. Bottle taking window, 39. Touch display screen, 40. Input module, 41. Display module, 42. Power module, 43. Sample placement area. Detailed implementation manners

[0051] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present application clearer, the present application will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application. The methods used in the present invention are all conventional methods unless otherwise specified; the raw materials and devices used are all conventional commercially available products unless otherwise specified.

[0052] Embodiment 1:

[0053] Preparation of the reagent kit:

[0054] 1) Preparation of Reagent A

[0055] Mix bovine serum albumin BSA, Tris buffer, coagulant, and preservative, and adjust the pH with NaOH solution and HCl solution to finally obtain Reagent A with a pH of 8.0 for standby; among them, Reagent A contains 2% bovine serum albumin BSA, 2.4 g / L Tris, 1 g / L PEG6000, and 0.5 g / L sodium azide.

[0056] 2) Preparation of Reagent B

[0057] ① Add NaOH solution and HCl solution to MES buffer to adjust the pH to obtain MES buffer with a pH of 6.0, where the concentration of MES buffer is 50 mM;

[0058] Take a part of the MES buffer with a pH of 6.0, add polystyrene microspheres with a diameter of 80 nm to it for washing, and after washing, centrifuge to discard the supernatant, and retain the precipitate I containing polystyrene microspheres;

[0059] ② Resuspend the precipitate I obtained in step ① in 50 mM MES buffer with a pH of 6.0, control the ultrasonic power at 60%, so that the concentration of polystyrene microspheres in the MES buffer is 2.5 g / L, add 2 g / L EDAC and 3 g / L NHS thereto to form a mixed solution I, activate it at room temperature for 20 min, centrifuge to remove the supernatant, and obtain precipitate II containing polystyrene microspheres for standby;

[0060] ③ Add NaOH solution and HCl solution to 30 mM HEPES buffer to adjust the pH to obtain HEPES buffer with a pH of 7.0; resuspend the precipitate II obtained in step ② in 30 mM HEPES buffer with a pH of 7.0, control the ultrasonic power at 60% to form a mixed solution II; add a coronavirus antibody 5A6 to the mixed solution II so that the concentration of the coronavirus antibody 5A6 in the mixed solution II is 0.5 mg / ml, oscillate at a temperature of 20 °C and a stirring speed of 100 r / min for 1 hour, centrifuge at 15000 r / min to remove the supernatant, and retain precipitate III containing polystyrene microspheres for standby;

[0061] ④ Add NaOH solution and HCl solution to PBS buffer to adjust the pH, and then add bovine serum albumin (BSA) thereto to obtain a mixed solution III with a pH of 7.4, so that the concentration of PBS buffer in the mixed solution III is 50 mM and the concentration of bovine serum albumin (BSA) in the mixed solution III is 20 g / L; resuspend the precipitate III obtained in step ③ in the mixed solution III, control the temperature at 25 °C, the ultrasonic power at 30%, and the stirring speed at 120 r / min, oscillate and seal overnight, centrifuge at 15000 r / min to remove the supernatant, and retain precipitate IV containing polystyrene microspheres;

[0062] ⑤ Add trehalose and glycerol to HEPES buffer to obtain a mixed solution IV so that the concentration of HEPES buffer in the mixed solution IV is 30 mM, the concentration of trehalose in the mixed solution IV is 35 g / L, and the concentration of glycerol in the mixed solution IV is 35 g / L; add the precipitate IV obtained in step ④ to the above-obtained mixed solution IV and resuspend it ultrasonically, control the ultrasonic power at 30 - 60% to obtain an immunomicrosphere mixed solution for detecting a coronavirus, that is, the preparation of reagent B is completed.

[0063] Detection on the instrument:

[0064] After mixing different concentrations of the sample to be tested (2 μL) and reagent A (150 μL), add immunocolloidal emulsion (reagent B, 50 μL), and measure the light intensity with a spectrometer after incubation for 3 min. AsFigure 1 As shown, when the sample to be tested is diluted to 4.35×10 -13 mol / L, the light intensity is stable and significantly different from the control, and the detection is completed within 5 minutes.

[0065] Example 2:

[0066] Preparation of the kit:

[0067] 1) Preparation of Reagent A

[0068] Mix bovine serum albumin BSA, Tris buffer, coagulant, and preservative, and adjust the pH with NaOH solution and HCl solution to finally obtain Reagent A with a pH of 8.0 for standby; among them, Reagent A contains 2% bovine serum albumin BSA, 2.4 g / L Tris, 5 g / L PEG 6000, and 5 g / L hydroxybenzoic acid.

[0069] 2) Preparation of Reagent B

[0070] ① Add NaOH solution and HCl solution to MES buffer to adjust the pH to obtain MES buffer with a pH of 6.0, where the concentration of MES buffer is 50 mM;

[0071] Take a part of the MES buffer with a pH of 6.0, add polystyrene microspheres with a diameter of 200 nm for washing, and after washing, centrifuge to discard the supernatant, and retain the precipitate I containing polystyrene microspheres;

[0072] ② Resuspend the precipitate I obtained in step ① ultrasonically in 50 mM MES buffer with a pH of 6.0, control the ultrasonic power at 60%, so that the concentration of polystyrene microspheres in the MES buffer is 2.5 g / L, add 20 g / L EDAC and 30 g / L NHS to form a mixed solution I, activate at room temperature for 30 minutes, centrifuge to remove the supernatant, and obtain the precipitate II containing polystyrene microspheres for standby;

[0073] ③ Add NaOH solution and HCl solution to 30 mM HEPES buffer to adjust the pH to obtain HEPES buffer with a pH of 7.0; resuspend the precipitate II obtained in step ② ultrasonically in 30 mM HEPES buffer with a pH of 7.0, control the ultrasonic power at 60% to form a mixed solution II; add a coronavirus antibody 5A6 to the mixed solution II so that the concentration of the coronavirus antibody 5A6 in the mixed solution II is 3 mg / ml, shake at a temperature of 30°C and a stirring speed of 300 r / min for 3 hours, centrifuge at 15000 r / min to remove the supernatant, and retain the precipitate III containing polystyrene microspheres for standby;

[0074] ④ Add NaOH solution and HCl solution to the PBS buffer to adjust the pH, and then add bovine serum albumin (BSA) to it to obtain the mixed solution III with a pH of 7.4, so that the concentration of the PBS buffer in the mixed solution III is 50 mM and the concentration of bovine serum albumin (BSA) in the mixed solution III is 20 g / L; Resuspend the precipitate III obtained in step ③ in the mixed solution III by ultrasonic treatment, where the temperature is controlled at 25 °C, the ultrasonic power is controlled at 60%, the stirring speed is controlled at 200 r / min, oscillate and seal overnight, and centrifuge at 15000 r / min to remove the supernatant, and retain the precipitate IV containing polystyrene microspheres;

[0075] ⑤ Add trehalose and glycerol to the HEPES buffer to obtain the mixed solution IV so that the concentration of the HEPES buffer in the mixed solution IV is 30 mM, the concentration of trehalose in the mixed solution IV is 35 g / L, and the concentration of glycerol in the mixed solution IV is 35 g / L; Add the precipitate IV obtained in step ④ into the above-obtained mixed solution IV and resuspend it by ultrasonic treatment, and control the ultrasonic power at 30 - 60% to obtain the immune microsphere mixed solution for detecting a coronavirus, that is, the preparation of reagent B is completed.

[0076] Detection on the machine:

[0077] Mix different concentrations of the sample to be tested (2 μL) with reagent A (150 μL), then add the immunocolloidal emulsion (reagent B, 50 μL), and measure the light intensity with a spectrometer after incubating for 3 min. As Figure 2 shown, when the sample to be tested is diluted to 4.35×10 -13 mol / L, the light intensity is stable and significantly different from the control, and the detection is completed within 5 min.

[0078] Example 3:

[0079] Preparation of the kit:

[0080] 1) Preparation of reagent A

[0081] Mix bovine serum albumin BSA, Tris buffer, coagulant, and preservative, and adjust the pH with NaOH solution and HCl solution to finally obtain reagent A with a pH of 8.0 for standby; Among them, reagent A contains 2% bovine serum albumin BSA, 2.4 g / L Tris, 3 g / L PEG 8000, and 2 g / L hydroxybenzoic acid.

[0082] 2) Preparation of reagent B

[0083] ① Add NaOH solution and HCl solution to the MES buffer to adjust the pH to obtain the MES buffer with a pH of 6.0, where the concentration of the MES buffer is 50 mM;

[0084] Take a portion of MES buffer with a pH of 6.0, add polystyrene microspheres with a diameter of 300 nm to it for washing. After completion of washing, centrifuge and discard the supernatant, and retain the precipitate I containing polystyrene microspheres.

[0085] ② Resuspend the precipitate I obtained in step ① ultrasonically in 50 mM MES buffer with a pH of 6.0, control the ultrasonic power at 60%, so that the concentration of polystyrene microspheres in the MES buffer is 2.5 g / L. Add 20 g / L EDAC and 30 g / L NHS to it to form a mixed solution I, activate at room temperature for 40 min, centrifuge to remove the supernatant, and obtain the precipitate II containing polystyrene microspheres for standby.

[0086] ③ Add NaOH solution and HCl solution to 30 mM HEPES buffer to adjust the pH to obtain HEPES buffer with a pH of 7.0; Resuspend the precipitate II obtained in step ② ultrasonically in 30 mM HEPES buffer with a pH of 7.0, control the ultrasonic power at 60% to form a mixed solution II; Add a coronavirus antibody 5A6 to the mixed solution II so that the concentration of the coronavirus antibody 5A6 in the mixed solution II is 3 mg / ml. At a temperature of 30 °C and a stirring speed of 300 r / min, oscillate for 4 hours, centrifuge at 15000 r / min to remove the supernatant, and retain the precipitate III containing polystyrene microspheres for standby.

[0087] ④ Add NaOH solution and HCl solution to PBS buffer to adjust the pH, and then add bovine serum albumin (BSA) to obtain a mixed solution III with a pH of 7.4, so that the concentration of PBS buffer in the mixed solution III is 50 mM and the concentration of bovine serum albumin (BSA) in the mixed solution III is 20 g / L; Resuspend the precipitate III obtained in step ③ ultrasonically in the mixed solution III, control the temperature at 25 °C, the ultrasonic power at 60%, and the stirring speed at 300 r / min, oscillate and seal overnight, centrifuge at 15000 r / min to remove the supernatant, and retain the precipitate IV containing polystyrene microspheres.

[0088] ⑤ Add trehalose and glycerol to HEPES buffer to obtain a mixed solution IV so that the concentration of HEPES buffer in the mixed solution IV is 30 mM, the concentration of trehalose in the mixed solution IV is 35 g / L, and the concentration of glycerol in the mixed solution IV is 35 g / L; Add the precipitate IV obtained in step ④ to the above-obtained mixed solution IV, ultrasonically resuspend it, and control the ultrasonic power at 30 - 60% to obtain an immunomicrosphere mixed solution for detecting a coronavirus, that is, the preparation of reagent B is completed.

[0089] Instrument detection:

[0090] After mixing different concentrations of the sample to be measured (3 μL) with Reagent A (150 μL), Immune Colloidal Emulsion (Reagent B, 50 μL) was added. After incubation for 3 min, the light intensity was measured using a spectrometer. As Figure 3 shown, when the sample to be measured was diluted to 4.35×10 -13 mol / L, the light intensity was stable and significantly different from the control, and the detection was completed within 5 min.

[0091] Example 4:

[0092] Preparation of the kit:

[0093] 1) Preparation of Reagent A

[0094] Bovine Serum Albumin BSA, Tris buffer, coagulant, and preservative were mixed, and the pH was adjusted with NaOH solution and HCl solution to finally obtain Reagent A with a pH of 8.0 for standby; among them, Reagent A contains 2% Bovine Serum Albumin BSA, 2.4 g / L Tris, 5 g / L PEG 8000, and 4 g / L hydroxybenzoic acid.

[0095] 2) Preparation of Reagent B

[0096] ① NaOH solution and HCl solution were added to the MES buffer to adjust the pH to obtain a MES buffer with a pH of 6.0, and the concentration of the MES buffer was 50 mM;

[0097] Take a part of the MES buffer with a pH of 6.0, add polystyrene microspheres with a diameter of 160 nm for washing, and after washing, centrifuge to discard the supernatant, and retain the precipitate I containing polystyrene microspheres;

[0098] ② The precipitate I obtained in step ① was ultrasonically resuspended in 50 mM MES buffer with a pH of 6.0, and the ultrasonic power was controlled at 60% to make the concentration of polystyrene microspheres in the MES buffer 2.5 g / L. 15 g / L EDAC and 20 g / L NHS were added thereto to form a mixed solution I, which was activated at room temperature for 40 min, and the supernatant was removed by centrifugation to obtain a precipitate II containing polystyrene microspheres for standby;

[0099] ③ Add NaOH solution and HCl solution to 30 mM HEPES buffer to adjust the pH and obtain HEPES buffer with a pH of 7.0; ultrasonically resuspend the precipitate II obtained in step ② in 30 mM HEPES buffer with a pH of 7.0, control the ultrasonic power at 60%, and form mixed solution II; add a coronavirus antibody 5A6 to mixed solution II to make the concentration of the coronavirus antibody 5A6 in mixed solution II 2.0 mg / ml, oscillate at a temperature of 30 °C and a stirring speed of 200 r / min for 3 hours, centrifuge at 15000 r / min to remove the supernatant, and retain the obtained precipitate III containing polystyrene microspheres for standby;

[0100] ④ Add NaOH solution and HCl solution to PBS buffer to adjust the pH, and then add bovine serum albumin (BSA) to obtain mixed solution III with a pH of 7.4, make the concentration of PBS buffer in mixed solution III 50 mM, and the concentration of bovine serum albumin (BSA) in mixed solution III 20 g / L; ultrasonically resuspend the precipitate III obtained in step ③ in mixed solution III, control the temperature at 25 °C, the ultrasonic power at 50%, and the stirring speed at 250 r / min, oscillate and seal overnight, centrifuge at 15000 r / min to remove the supernatant, and retain the obtained precipitate IV containing polystyrene microspheres;

[0101] ⑤ Add trehalose and glycerol to HEPES buffer to obtain mixed solution IV, make the concentration of HEPES buffer in mixed solution IV 30 mM, the concentration of trehalose in mixed solution IV 35 g / L, and the concentration of glycerol in mixed solution IV 35 g / L; add the precipitate IV obtained in step ④ to the above-obtained mixed solution IV and ultrasonically resuspend it, control the ultrasonic power at 30 - 60%, and obtain an immunomicrosphere mixed solution for detecting a coronavirus, that is, complete the preparation of reagent B.

[0102] Detection on the machine:

[0103] After mixing different concentrations of the sample to be measured (4 μL) and reagent A (150 μL), add immunocolloid emulsion (reagent B, 50 μL), and measure the light intensity with a spectrometer after incubation for 3 min. As Figure 4 shown, when the sample to be measured is diluted to 4.35×10 -13 mol / L, the light intensity is stable and significantly different from the control, and the detection is completed within 5 min.

[0104] Example 5:

[0105] Preparation of the kit:

[0106] 1) Preparation of reagent A

[0107] Mix bovine serum albumin (BSA), Tris buffer, coagulant, and preservative, and adjust the pH with NaOH solution and HCl solution to finally obtain reagent A with a pH of 8.0 for standby; among them, reagent A contains 2% bovine serum albumin (BSA), 2.4 g / L Tris, 2 g / L PEG 6000, and 5 g / L sodium azide.

[0108] 2) Prepare reagent B

[0109] ① Add NaOH solution and HCl solution to the MES buffer to adjust the pH to obtain a MES buffer with a pH of 6.0, where the concentration of the MES buffer is 50 mM;

[0110] Take a part of the MES buffer with a pH of 6.0, add polystyrene microspheres with a diameter of 300 nm to it for washing. After washing, centrifuge and discard the supernatant to obtain precipitate I containing polystyrene microspheres;

[0111] ② Resuspend precipitate I obtained in step ① in 50 mM MES buffer with a pH of 6.0 by ultrasonic treatment, control the ultrasonic power at 60%, so that the concentration of polystyrene microspheres in the MES buffer is 2.5 g / L, add 10 g / L EDAC and 20 g / L NHS to it to form mixed solution I, activate it at room temperature for 40 min, centrifuge to remove the supernatant, and obtain precipitate II containing polystyrene microspheres for standby;

[0112] ③ Add NaOH solution and HCl solution to 30 mM HEPES buffer to adjust the pH to obtain a HEPES buffer with a pH of 7.0; resuspend precipitate II obtained in step ② in 30 mM HEPES buffer with a pH of 7.0 by ultrasonic treatment, control the ultrasonic power at 60% to form mixed solution II; add a coronavirus antibody 5A6 to mixed solution II so that the concentration of the coronavirus antibody 5A6 in mixed solution II is 0.5 mg / ml, oscillate at a temperature of 250 °C and a stirring speed of 300 r / min for 1 hour, centrifuge at 15000 r / min to remove the supernatant, and obtain precipitate III containing polystyrene microspheres for standby;

[0113] It should be noted that in step 15, the temperature of 250 °C seems rather unusual. Please double-check if this is the correct value. If it's a misprint, it may need to be corrected according to the actual situation.④ Add NaOH solution and HCl solution to the PBS buffer to adjust the pH, and then add bovine serum albumin (BSA) to it to obtain mixed solution III with a pH of 7.4, such that the concentration of the PBS buffer in the mixed solution III is 50 mM and the concentration of bovine serum albumin (BSA) in the mixed solution III is 20 g / L; ultrasonically resuspend the precipitate III obtained in step ③ in the mixed solution III, with the temperature controlled at 25 °C, the ultrasonic power controlled at 40%, the stirring speed controlled at 200 r / min, oscillate and seal overnight, centrifuge at 15000 r / min to remove the supernatant, and retain the precipitate IV containing polystyrene microspheres;

[0114] ⑤ Add trehalose and glycerol to the HEPES buffer to obtain mixed solution IV such that the concentration of the HEPES buffer in the mixed solution IV is 30 mM, the concentration of trehalose in the mixed solution IV is 35 g / L, and the concentration of glycerol in the mixed solution IV is 35 g / L; add the precipitate IV obtained in step ④ into the above-obtained mixed solution IV and ultrasonically resuspend it, with the ultrasonic power controlled at 30 - 60% to obtain an immunomicrosphere mixed solution for detecting a coronavirus, thus completing the preparation of reagent B.

[0115] Machine detection:

[0116] Mix different concentrations of the sample to be measured (5 μL) and reagent A (150 μL), then add the immunocolloidal emulsion (reagent B, 50 μL), and measure the light intensity with a spectrometer after incubating for 3 min. As Figure 5 shown, when the sample to be measured is diluted to 4.35×10 -14 mol / L, the light intensity is stable but there is no significant difference from the control; when the sample to be measured is diluted to 4.35×10 -13 mol / L, the light intensity is stable and there is a significant difference from the control, and the detection is completed within 5 min.

[0117] As Figure 10 , Figure 11 shown, for the machine detection equipment used in the above Examples 1 - 5 as a rapid detection equipment for coronavirus, it is provided with a light source 1, a slit 2, a grating 3, a detection mechanism, and a spectrometer 4. The detection mechanism is provided with a sample bottle 5 for containing the liquid reagent to be measured; the light source 1 outputs a stable beam with a fixed wavelength. The stable beam enters the grating 3 after passing through the slit of the slit 2, is split by the grating 3 and then passes through the sample bottle 5 and the liquid reagent to be measured inside it to form a transmitted beam, and the transmitted beam enters the spectrometer 4 for detecting the light intensity.

[0118] The wavelength of the light source 1 is a fixed wavelength, and the wavelength range is preferably 400 - 1000 nm. A beam within this wavelength range can produce a more stable transmitted beam after passing through the sample bottle 5 and the liquid reagent to be measured inside it. The light source 1 is preferably a tungsten halogen light source.

[0119] The purpose of the slit 2 is to reduce the beam intensity of the light source 1 so that the intensity of the transmitted beam is within the working range of the spectrometer 4. It is preferably an adjustable slit with an adjustment range of 0 - 5 mm. For light sources 1 with different intensities, the same or corresponding beam intensity can be obtained by adjusting the slit width of the slit 2. The slit 2 is preferably a symmetric slit with bilateral adjustability.

[0120] The purpose of the grating 3 is to disperse the beam transmitted through the slit 2 to obtain a highly pure monochromatic beam. According to the Lambert - Beer law, a highly pure monochromatic beam can improve the detection accuracy of the spectrometer 4. The grating 3 is preferably a transmission grating.

[0121] The sample vial 5 is used to hold the liquid reagent to be tested. The liquid reagent to be tested contains antibodies corresponding to the types of viruses to be detected. For example, when detecting the COVID - 19 virus, the liquid reagent to be tested is the corresponding antibody of this virus, such as the 5A6 and / or 8A13 antibodies. To make the beam more easily penetrate the vial body 28 of the sample vial 5 and minimize the impact on the beam intensity, the vial body 28 of the sample vial 5 is preferably made of a colorless and transparent material, such as colorless and transparent glass, etc.; the beam preferably penetrates vertically into the vial body 28 of the sample vial 5 and then exits the vial body 28 vertically. Since the liquid reagent to be tested contains viruses, to prevent the liquid reagent to be tested from contaminating the external environment or being contaminated by the external environment, after the liquid reagent to be tested is filled into the sample vial 5, it is preferably sealed in time. According to the actual situation of virus detection, the volume of the liquid reagent to be tested is usually 200 - 700 μL, so the internal cavity volume of the sample vial 5 is less than 1 mL.

[0122] The spectrometer 4 is used to detect the intensity of the transmitted beam formed by passing through the sample vial 5 and the liquid reagent to be tested inside it. By detecting the intensity of the transmitted beam, the number of viruses contained in the sampling sample added to the sample vial 5 can be determined. The virus (i.e., the sample to be tested) specifically binds to the liquid reagent to be tested (i.e., reagent A and reagent B), affecting the turbidity of the liquid reagent to be tested, and the turbidity of the liquid reagent to be tested is positively correlated with the number of viruses, that is, the more viruses, the higher the turbidity of the liquid reagent to be tested; there is a correlation between the turbidity of the liquid reagent to be tested and the intensity value of the transmitted beam detected by the spectrometer 4, that is, as the turbidity of the liquid reagent to be tested increases, the intensity of the transmitted beam formed by the same - intensity beam passing through the sample vial 5 and the liquid reagent to be tested inside it becomes lower; by detecting the intensity value of the transmitted beam with the spectrometer 4 and looking up the corresponding value on the virus number - light intensity value standard curve for the detected beam intensity value, the number of viruses can be obtained, and the sample detection is completed. The spectrometer 4 is usually an infrared spectrometer.

[0123] The method for making the standard curve of virus quantity - light intensity value is as follows: The same liquid reagent to be measured is equally divided and filled into multiple identical sample bottles 5; Different determined quantities of viruses are respectively added into the liquid reagent to be measured in the sample bottles 5, and mixed evenly to prepare the liquid reagent to be measured containing virus standards with different concentrations. Then the sample bottles 5 are sealed; The above sample bottles 5 are respectively detected by using the device of the present invention. During the operation process, the difference is only that the concentrations of the virus standards in the liquid reagent to be measured in the sample bottles 5 are different, and the others are the same. The corresponding light intensity values are respectively obtained and fitted into the standard curve of virus quantity - light intensity value. Figure 6 It is the standard curve of virus quantity - light intensity value in Example 2.

[0124] The working principle of the rapid coronavirus detection device of the present invention is: The present invention provides a rapid coronavirus detection device, which does not rely on the existing technical professional detection sites and can be installed in public places such as airports and stations for on - site rapid virus detection. Detection can be carried out on - site, greatly shortening the detection process, so as to meet the diagnosis needs of a large number of suspected infected persons and quarantined observers. The on - site rapid virus detection device is provided with a light source 1. The stable light beam with a fixed wavelength generated by the light source 1 passes through the slit 2 to obtain a light beam with appropriate intensity, and then passes through the grating 3 to obtain a highly pure monochromatic light beam. The monochromatic light beam passes through the sample bottle 5 and the liquid reagent to be measured inside it to generate a transmitted light beam. The transmitted light beam enters the spectrometer 4 and can be converted into an intensity signal, and the intensity value of the transmitted light beam detected by the spectrometer 4 is obtained. The turbidity of the liquid reagent to be measured in the sample bottle 5 is positively correlated with the quantity of the virus, and there is a correlation between the turbidity of the liquid reagent to be measured and the intensity value of the transmitted light beam detected by the spectrometer 4, that is, as the turbidity of the liquid reagent to be measured increases, the intensity of the transmitted light beam formed by the same - intensity light beam passing through the sample bottle 5 and the liquid reagent to be measured inside it is lower; By detecting the intensity value of the transmitted light beam by the spectrometer 4, looking up the corresponding value in the standard curve of virus quantity - light intensity value for the detected light beam intensity value, the virus quantity can be obtained, and the virus detection of the liquid reagent to be measured is completed.

[0125] In some embodiments, the detection mechanism of the present invention preferably further includes a platform base 6 for placing and fixing the sample bottle 5 to make the state of the sample bottle 5 stable and reliable during the detection process. The platform base 6 can be an existing device or Figure 10 、 Figures 12 - 14The platform base 6 shown, for the convenience of installation and removal of the sample vial 5, is provided with a C-shaped notch 20 for placing the sample vial 5. When placing the sample vial 5, the bottom of the sample vial 5 is inserted horizontally from the opening of the C-shaped notch 20, so that the bottom of the sample vial 5 is snap-fitted inside the C-shaped notch 20, thus completing the placement and fixation of the sample vial 5. As a further preference, a first installation blind hole 25 adapted to the bottom of the sample vial 5 is provided inside the C-shaped notch 20, and the bottom wall from the bottom wall of the first installation blind hole 25 to the bottom wall of the opening of the C-shaped notch 20 shows a slow upward trend, which is convenient for the bottom of the sample vial 5 to enter the inside of the C-shaped notch 20 horizontally from the opening of the C-shaped notch 20, and then be inserted into the first installation blind hole 25, making the placement of the sample vial 5 more fixed and reliable. After the virus detection is completed, the sample vial 5 is removed by an operation opposite to the installation sequence, which is convenient and fast. In addition, the platform base 6 can also be Figure 15 the platform base 6 shown. The platform base 6 is provided with a second installation blind hole 26, and an elastic piece 27 is connected to the side wall of the second installation blind hole 26. The number of the elastic pieces 27 is multiple, and they are evenly distributed on the side wall of the second installation blind hole 26 to form a biting structure with a hole in the middle. When installing the sample vial 5, the bottom of the sample vial 5 is inserted into the second installation blind hole 26 from top to bottom, and the elastic pieces 27 forming the biting structure are deformed and snap-fitted on the outer wall of the bottom of the sample vial 5, completing the installation of the sample vial 5. After the virus detection is completed, the sample vial 5 is removed by an operation opposite to the installation sequence, and the elastic pieces 27 forming the biting structure return to their original state, which is convenient and fast.

[0126] The sample vial 5 can use commercially available products, or can also use such as Figure 11 the sample vial 5 shown. The bottle body 28 of the sample vial 5 is of a cylindrical structure. The upper part of the bottle body 28 is provided with a bottle mouth, and the bottle mouth is threadedly sealed with a bottle cap 29. The periphery of the bottom of the bottle body 28 is provided with a rounded corner structure 30; a cylindrical cavity communicating with the bottle mouth is provided inside the sample vial 5, and the cylindrical cavity is coaxially arranged with the bottle body 28 of the sample vial 5.

[0127] In some embodiments, by Figure 9 、 Figure 10As shown, the detection mechanism preferably further includes a stepping motor 7 and a transmission device. The transmission device includes an annular rack 8, an annular bracket 9, a support arm 10, and a gear 11. The annular rack 8 is coaxially sleeved inside the annular bracket 9. The teeth of the annular rack 8 are arranged on the inner ring of the annular rack 8. One end of the support arm 10 is connected to the annular rack 8. The other end of the support arm 10 faces the annular bracket 9 and extends out. The body of the support arm 10 is slidably connected to the annular bracket 9. The power output shaft of the stepping motor 7 is connected to the gear 11. The gear 11 meshes with the teeth on the inner ring of the annular rack 8. The number of support arms 10 is set to be multiple, and the multiple support arms 10 are spaced apart on the annular rack 8, preferably distributed in a circular array centered on the central axis of the annular rack 8. A platform base 6 is connected to each support arm 10. Under the power output of the stepping motor 7, through the transmission of the gear 11, the annular rack 8 is driven to rotate, thereby driving the support arm 10 together with the platform base 6 on the support arm 10 to rotate around the central axis of the annular rack 8. When the sample bottle 5 on one platform base 6 completes the virus detection, driven by the stepping motor 7, the platform base 6 together with the sample bottle 5 is moved out of the detection point. At the same time, the next platform base 6 together with the sample bottle 5 to be tested for virus is moved into the detection point, realizing automatic sample injection and sampling, which is more convenient and fast.

[0128] In some embodiments, by Figure 10 As shown, in order to make the driving of the annular rack 8 more stable and reliable during the rotation process, a slider 12 is preferably connected to the body of the support arm 10, and the annular bracket 9 is provided with a circumferential slideway 13 matching the slider 12.

[0129] In some embodiments, by Figure 9 、 Figure 10 As shown, in order to further ensure that the liquid reagent to be tested in the sample bottle 5 is in a constant temperature state and make the virus detection result more accurate, a constant temperature component is preferably further provided on the outer side of the annular bracket 9. The constant temperature component includes a heat sink 14 and a semiconductor refrigeration chip 15 connected to each other. The other end of the support arm 10 faces the annular bracket 9 and extends above the constant temperature component. The platform base 6 is arranged above the heat sink 14. The heat sink 14 provides heat for the liquid reagent to be tested in the sample bottle 5 installed on the platform base 6. The present invention uses the semiconductor refrigeration chip 15 to adjust the temperature of the heat sink 14 to a constant temperature. The semiconductor refrigeration chip 15 has two sides. One side has an endothermic function. After absorbing heat, the temperature decreases, realizing a cooling effect. The other side has a heat dissipation function, realizing a heating effect.

[0130] In some embodiments, by Figure 10As shown, in order to reduce labor costs and further improve the mechanization level of the present invention, the detection mechanism preferably further comprises a pushing device. The pushing device is provided with a baffle and a leak plate 16. The baffle is provided with a supporting portion 17 and a blocking and pushing portion 18 which are connected to each other. The leak plate 16 is provided with leak holes 19 which are arranged on one side of the baffle. The supporting portion 17 is connected to the leak plate 16, and the blocking and pushing portion 18 is arranged above the platform base 6 and is used for pushing down the sample bottle 5 that has completed the detection and then entering the leak holes 19. After the virus detection of the sample bottle 5 on a platform base 6 is completed, it is driven by the power output of the stepping motor 7, transmitted by the gear 11, and drives the annular rack 8 to rotate self-driven, so as to drive the platform base 6 together with the sample bottle 5 to move out of the detection point. The bottle body 28 of the sample bottle 5 stops moving after hitting the blocking and pushing portion 18 after being moved out, while the platform base 6 at the lower part of the sample bottle 5 still rotates together with the supporting arm 10, so that the sample bottle 5 is pushed down by the blocking and pushing portion 18 and falls into the leak holes 19, completing the removal work of the sample bottle 5 on the platform base 6. A waste sample box 31 is preferably placed directly below the leak holes 19. The waste sample box 31 is a box-shaped structure with an open upper part. After the sample bottle 5 is pushed down, it falls through the leak holes 19 to the waste sample box 31 as shown in Figure 7 shown.

[0131] In some embodiments, in order to make the removal work of the above-mentioned sample bottle 5 on the platform base 6 smoother, it is preferably to use a platform base 6 as shown in Figures 12 - 14 shown, and the leak holes 19 are arranged at intervals on the opening side of the C-shaped notch 20. The blocking and pushing portion 18 is provided with a V-shaped opening 21, and the orientation of the V-shaped opening 21 is the same as that of the C-shaped notch 20, as shown in Figure 10 shown. After the sample bottle 5 is pushed down by the blocking and pushing portion 18, it topples and falls into the leak holes 19 in the opening direction of the C-shaped notch 20. The setting of the V-shaped opening 21 of the blocking and pushing portion 18, on the one hand, increases the contact area with the sample bottle 5 and reduces the pressure at the moment when the sample bottle 5 touches the blocking and pushing portion 18; on the other hand, after the sample bottle 5 is pushed down, the two side edges of the V-shaped opening 21 can limit the toppling direction of the sample bottle 5, so that the sample bottle 5 smoothly topples and falls into the leak holes 19 and falls through the leak holes 19 to the waste sample box 31 as shown in Figure 7 shown below it.

[0132] In some embodiments, as shown in Figure 10 shown, the supporting portion 17 and the blocking and pushing portion 18 are preferably plate-shaped structures respectively. The supporting portion 17 and the blocking and pushing portion 18 are connected to form an inverted L-shaped structure. Among them, the supporting portion 17 is arranged at the outermost end far from the supporting arm 10, and the blocking and pushing portion 18 is arranged at intervals above the platform base 6, so that the annular rack 8 drives the supporting arm 10 together with the platform base 6 to rotate without obstruction.

[0133] In some embodiments, as shown in Figure 11As shown, a rounded corner structure 30 is preferably provided around the bottom of the bottle body 28. When the sample bottle 5 is pushed down, on the one hand, it makes the sample bottle 5 easier to pour; on the other hand, it protects the periphery of the bottom of the sample bottle 5 from damage.

[0134] In some embodiments, by Figure 10 、 Figures 16 - 18 As shown, the detection mechanism is also preferably provided with a detection cover 32. The detection cover 32 serves as the above-mentioned detection point and is arranged between the grating 3 and the spectrometer 4. The bottom surface of the detection cover 32 is provided with a through groove opening downward, making the whole detection cover 32 in an n-shaped structure. The through groove serves as the channel for the sample bottle 5 and its platform base 6 to enter and exit. The through groove includes a connected sample bottle channel 33 and a platform base channel 34. One light beam through hole 35 connected to the sample bottle channel 33 is respectively opened on the left and right side walls of the sample bottle channel 33, and the two light beam through holes 35 are arranged opposite to each other; the platform base channel 34 is arranged below the sample bottle channel 33, and the width of the platform base channel 34 is greater than the width of the sample bottle channel 33. After the platform base 6 of the sample bottle 5 is placed into the platform base channel 34, a detection space is formed with the detection cover 32, and the sample bottle 5 is arranged in this detection space. As Figure 17 As shown, the sample bottle channel 33 and the platform base channel 34 are arc-shaped, which is consistent with the running trajectories of the sample bottle 5 and the platform base 6. Driven by the power output of the stepping motor 7, through the transmission of the gear 11, the annular rack 8 rotates self-driven, thereby driving the support arm 10 together with the platform base 6 on the support arm 10 to rotate around the central axis of the annular rack 8, driving the sample bottle 5 of one platform base 6 into the through groove. The sample bottle 5 stops at the position where the two light beam through holes 35 are arranged opposite to each other. At this time, the stable light beam with a fixed wavelength generated by the light source 1 passes through the slit 2 to obtain a light beam with appropriate intensity, and then passes through the grating 3 to obtain a highly pure monochromatic light beam. The monochromatic light beam enters one light beam through hole 35, and then generates a transmitted light beam after passing through the sample bottle 5 and the liquid reagent to be measured inside it. The transmitted light beam enters the spectrometer 4 through the other light beam through hole 35 and can be converted into an intensity signal, and the intensity value of the transmitted light beam detected by the spectrometer 4 is obtained; after the virus detection is completed, driven by the stepping motor 7, the platform base 6 together with the sample bottle 5 is moved out of the detection cover 32. At the same time, the next platform base 6 together with the sample bottle 5 to be tested for virus is moved into the detection cover 32 for virus detection, realizing automatic sample injection and ejection, which is more convenient and fast. By Figure 10 As shown, during the process of the annular rack 8 driving the support arm 10 together with the platform base 6 on the support arm 10 to rotate around the central axis of the annular rack 8, the support arm 10 together with the platform base 6 on the support arm 10, the light source 1, the slit 2, the grating 3, and the detection cover 32 are all arranged at intervals above the support arm 10, making the rotation of the annular rack 8 driving the support arm 10 unobstructed and proceeding smoothly.

[0135] In some embodiments, by Figure 7 、Figure 8 As shown, the detection mechanism preferably further includes a housing 36, which is in a box structure. The light source 1, slit 2, grating 3, detection mechanism, and spectrometer 4 are arranged inside the housing 36 to further ensure that virus detection is not affected by the external environment. A bottle loading window 37 and a bottle taking window 38 are provided on the side wall of the housing 36. The bottle loading window 37 is directly above the bottle taking window 38, and the leak plate 16 separates the bottle loading window 37 and the bottle taking window 38. The bottle loading window 37 is used to place the sample bottle 5 of the virus to be detected into the platform base 6, and a waste sample box 31 is placed in the bottle taking window 38. The waste sample box 31 is arranged directly below the leakage hole 19. A touch display screen 39 is also connected to the surface of the housing 36, which is mainly used to display the detection results of the virus in the liquid reagent to be detected.

[0136] In some embodiments, Figure 9 、 Figure 19 As shown, to further improve the automation level of the present invention, the on-site rapid virus detection device preferably further includes an intelligent control system, an infrared sensor 22, and a temperature sensor 23. The infrared sensor 22 is arranged on one side of the platform base 6 to detect whether the sample bottle 5 is placed on the platform base 6; the temperature sensor 23 is connected to the heat sink 14; the intelligent control system is provided with an intelligent control device 24, and the intelligent control device 24 can generally use a single-chip microcomputer; the intelligent control device 24 is connected to the infrared sensor 22, temperature sensor 23, stepping motor 7, and semiconductor refrigeration sheet 15 through control lines respectively.

[0137] When the sample bottle 5 is placed on the detection platform base 6, the infrared sensor 22 generates a high-level electrical signal and transmits it to the intelligent control device 24 through the control line. After receiving the high-level electrical signal, the internal timer of the intelligent control device 24 starts timing and sends an instruction to the stepping motor 7 through the control line. After receiving the instruction, the stepping motor 7 drives the annular rack 8 to rotate self. The rotation speed depends on the distance of the sample bottle 5 from the detection point and the set time. After the set time, which is the time required for the reagent chemical reaction, the internal timer sends a feedback signal to the intelligent control device 24. The intelligent control device 24 sends an instruction to the stepping motor 7 through the control line. After receiving the instruction, the stepping motor 7 stops driving, and the sample bottle 5 reaches the detection point for detection. The spectrometer 4 detects the intensity value of the transmitted light beam and transmits the signal to the intelligent control device 24 through the control line. After obtaining the signal generated by the spectrometer 4, the intelligent control device 24 calculates the number of viruses in the liquid reagent to be detected in the sample bottle 5 according to the internal virus quantity - light intensity value standard curve, and completes the detection of the virus in the liquid reagent to be detected in the sample bottle 5. When there are multiple sample bottles 5, each sample bottle 5 reaches the detection point after the set time of the internal timer in sequence, and the detection of the virus in the liquid reagent to be detected in the sample bottle 5 is completed. The working process is as above and will not be elaborated here.

[0138] The temperature sensor 23 monitors the temperature of the heat sink 14 and transmits the temperature sensing signal to the intelligent control device 24 through the control line. After receiving the sensing signal, when the temperature of the heat sink 14 is higher or lower than the preset temperature, the intelligent control device 24 sends a refrigeration command to the semiconductor refrigeration chip 15 through the control line. After receiving the command, the semiconductor refrigeration chip 15 controls the cooling and heating intensity to achieve the purpose of controlling the temperature.

[0139] The present invention further includes an input module 40, a display module 41, and a power supply module 42. The intelligent control device 24 is connected to the input module 40, the display module 41, and the power supply module 42 through the control line respectively. The input module 40 includes a keyboard and a touch display screen 39. By inputting a control command on the keyboard or the touch display screen 39, the control command is sent into the intelligent control device 24, and the input of the information of the intelligent control device 24 realizes its corresponding output control respectively. The display module 41 includes a touch display screen 39, which is mainly used to display the detection results of the liquid reagent virus to be detected. The power supply module 42 is mainly used to supply power to components such as the intelligent control system, the infrared sensor 22, the temperature sensor 23, the stepping motor 7, the semiconductor refrigeration chip 15, the keyboard, and the touch display screen 39.

[0140] As a further preferred embodiment, Figure 8 , Figure 9 As shown, a sample placement area 43 is provided at the bottle placement window 37 on the side wall close to the outer shell 36. The sample placement area 43 can accommodate three platform bases 6. An infrared sensor 22 is respectively provided corresponding to each platform base 6. The infrared sensor 22 is installed at the bottom of the inner cavity of the outer shell 36 and is used to detect whether a sample bottle 5 is placed on the platform base 6 rotated into the sample placement area 43 respectively, and transmits the sensing signal to the intelligent control device 24 through the control line. According to the actual situation, a sample bottle 5 can be placed on any one of the three platform bases 6. The purpose of setting three platform bases 6 is to enhance the placement tolerance of the sample bottle 5, improve the placement efficiency of the sample bottle 5, and ensure that the sample bottle 5 can be easily placed into the platform base 6 during the transmission of the ring gear 8. A plurality of support arms 10 are arranged at intervals on the ring gear 8 and are distributed in a circular array centered on the central axis of the ring gear 8. Each support arm 10 is connected to a platform base 6, and the platform bases 6 are also distributed in a circular array centered on the central axis of the ring gear 8.

[0141] An infrared sensor 22 is installed on the side of each of the above three platform bases 6. When the sample bottle 5 is placed in the platform base 6, the infrared sensor 22 will generate a high-level electrical signal. The infrared sensor 22 generates a high-level electrical signal and transmits it to the intelligent control device 24 through the control circuit. After receiving the high-level electrical signal, the internal timer of the intelligent control device 24 starts timing and sends an instruction to the stepping motor 7 through the control circuit. After receiving the instruction, the stepping motor 7 drives the annular rack 8 to rotate by a set angle. If the platform base 6 is set to N (N>0 and is a positive integer), then the set angle value is 360 / N. After completing the rotation of the set angle, the intelligent control device 24 sends an instruction to stop rotating to the stepping motor 7 through the control circuit. After receiving the instruction, the stepping motor 7 stops rotating, and the sample bottle 5 at the detection point is subjected to virus detection. The set time for the stepping motor 7 to stop rotating is the same as the virus detection time of the sample bottle 5. After this set time arrives, the intelligent control device 24 sends an instruction to the stepping motor 7 through the control circuit. After receiving the instruction, the stepping motor 7 drives the annular rack 8 to rotate by the set angle value of 360 / N in the same direction as before. At this time, the sample bottle 5 that has completed the detection is rotated out of the detection point, and the adjacent sample bottle 5 to be detected is rotated into the detection point. According to the above process, the intelligent control device 24 sends an instruction to the stepping motor 7 through the control circuit at intervals of the above set time. After receiving the instruction, the stepping motor 7 drives the annular rack 8 to rotate by the set angle value of 360 / N in the same direction as before. During the period when the annular rack 8 stops rotating, the operator can take out the sample bottle 5 that has completed the virus detection from the platform base 6 and place the sample bottle 5 to be detected into the platform base 6 in the sample placement area 43. The time is preset in the internal timer of the intelligent control device 24. After reaching the set time, a feedback signal is sent to the intelligent control device 24 by the internal timer. After receiving the signal, the intelligent control device 24 sends a corresponding instruction to the stepping motor 7.

[0142] It should be noted that the entire light beam propagation process can be carried out in a dark room. Especially when the sample bottle 5 is at the detection point, it should be in a dark room state so that the transmitted light beam formed by passing through the sample bottle 5 and the liquid reagent to be measured inside it is not affected by the external environment. Between the light source 1 and the slit 2, between the slit 2 and the grating 3, between the grating 3 and one side of the bottle body 28 of the sample bottle 5, and between the other side of the bottle body 28 of the sample bottle 5 and the spectrometer 4 can also be respectively connected by optical fibers; the light beam incident from one side of the bottle body 28 of the sample bottle 5 passes through the sample bottle 5 and the liquid reagent to be measured inside it to form a transmitted light beam and enters the optical fiber on the other side of the bottle body 28 of the sample bottle 5; the above light beam generated by the light source 1 is transmitted in the air and is changed to be transmitted in the optical fiber, making the light beam transmission more stable and reliable, and further improving the accuracy of the detection data of the spectrometer 4. Preferably, the optical fibers on both sides of the bottle body 28 of the sample bottle 5 are symmetrically arranged opposite to the connection with the bottle body 28 of the sample bottle 5, and the connection line between the two intersects the central axis of the bottle body 28 of the sample bottle 5.

[0143] The present invention does not rely on existing technical professional detection sites and can be installed in public places such as airports and stations for rapid detection of the novel coronavirus. Detection can be carried out on-site, greatly shortening the detection process, thus meeting the diagnostic needs of a large number of suspected infected persons and quarantined observers, which is of great significance. This can not only improve the efficiency and accuracy of the prevention and control of the coronavirus, safeguard the lives and health of the public, but also reasonably allocate medical resources during the peak period of the epidemic to avoid resource shortages. It will also provide valuable experience and technical support for the detection and prevention and control of other infectious diseases, help to timely detect and control the source of infection, and reduce the risk of the spread of infectious diseases.

[0144] The above are only the preferred embodiments of the present application and are not intended to limit the present application. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for preparing a coronavirus rapid detection kit, characterized in that: The steps include: S1. Prepare reagent A Mixing bovine serum albumin, buffer I, a coagulant, and a preservative, and adjusting the pH to finally obtain a reagent A with a pH of 8.0; wherein the configured reagent A contains 2% bovine serum albumin by mass, 2.4 g / L buffer I, 1-5 g / L coagulant, and 0.5-5 g / L preservative; S2. Prepare reagent B ① Adjusting the pH of buffer II to obtain a buffer II solution with a pH of 6.0; taking a portion of the buffer II solution, adding polystyrene microspheres with a diameter of 80 to 300 nm thereto for washing, centrifuging and discarding the supernatant after washing, and retaining a precipitate I containing polystyrene microspheres for standby use; ② Take another part of the buffer solution II prepared in step ①, add the precipitate I obtained in step ① thereto, and resuspend by ultrasound to make the concentration of the polystyrene microspheres in the buffer solution II be a preset concentration; then add 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide thereto to form a mixed solution I, wherein the concentration of 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride is 2 g / L, and the concentration of N-hydroxysuccinimide is 3 g / L; activate the mixed solution I at room temperature, centrifuge and remove the supernatant to obtain a precipitate II containing polystyrene microspheres for standby use; ③ Adjust the pH of buffer III to obtain a buffer III solution with a pH of 7.0; resuspend the precipitate II obtained in step ② in the buffer III solution by ultrasonication to form a mixed solution II; add coronavirus antibodies to the mixed solution II so that the concentration of the coronavirus antibodies in the mixed solution II is 0.5-3 mg / ml, stir and oscillate fully, centrifuge to remove the supernatant, and retain the precipitate III containing polystyrene microspheres for standby use; ④ Adjusting the pH of buffer IV, and then adding bovine serum albumin thereto to obtain a mixed solution III with a pH of 7.4; ultrasonically resuspending the precipitate III obtained in step ③ in the mixed solution III, stirring and oscillating sufficiently, and centrifuging to remove the supernatant, and retaining the precipitate IV containing polystyrene microspheres for standby use; ⑤ Add trehalose and glycerol to buffer solution V to obtain a mixed solution IV; add the precipitate IV obtained in step ④ to the mixed solution IV, resuspend by ultrasonication, and finally obtain reagent B.

2. The method for preparing a coronavirus rapid detection kit according to claim 1, characterized in that: The buffer I, buffer II, buffer III, buffer IV and buffer V are respectively one of tris(hydroxymethylaminomethane)-hydrochloric acid buffer, 4-hydroxyethylpiperazineethanesulfonic acid buffer, phosphate buffer, acetate buffer, glycine buffer and 2-(N-morpholino)ethanesulfonic acid buffer.

3. The method for preparing a coronavirus rapid detection kit according to claim 1, characterized in that: The coagulant includes one of PEG 6000 and PEG 8000.

4. The method for preparing a coronavirus rapid detection kit according to claim 1, characterized in that: The preservative includes one of sodium azide, sodium thimerosal, phenol, ethylparaben, and hydroxybenzoic acid.

5. The method for preparing a coronavirus rapid detection kit according to claim 1, characterized in that: In the step ②, the precipitate I is ultrasonically resuspended in the buffer solution II, and the ultrasonic power is controlled at 60%; the mixed solution I is activated at room temperature for 20 to 40 minutes.

6. The method for preparing a coronavirus rapid detection kit according to claim 1, characterized in that: In the step ③, the precipitate II is ultrasonically resuspended in the buffer solution III at a temperature of 20° C. and an ultrasonic power of 60%; the stirring speed is 100 to 300 r / min, the oscillation is 1 to 4 hours, and the centrifugal speed is 15000 r / min.

7. The method for preparing a coronavirus rapid detection kit according to claim 1, characterized in that: In the step ④, the precipitate III is ultrasonically resuspended in the mixed solution III, wherein the temperature is controlled at 25° C., the ultrasonic power is controlled at 30% to 60%, the stirring speed is controlled at 120 to 300 r / min, the mixture is oscillated overnight, and the centrifugal speed is 15000 r / min.

8. The method for preparing a coronavirus rapid detection kit according to claim 1, characterized in that: In the step ⑤, the precipitate IV is resuspended in the mixed solution IV by ultrasonication, and the ultrasonic power is controlled at 30% to 60%.

9. A method for preparing a coronavirus rapid detection kit according to any one of claims 1 to 8, characterized in that: In the step ①, the concentration of buffer II in the buffer II solution is 50 mM; In step ②, the concentration of polystyrene microspheres in the buffer II solution is 2.5 g / L; In step ③, the concentration of buffer III in the buffer III solution is 30 mM; In step ④, in the mixed solution III, the concentration of the buffer IV is 50 mM, and the concentration of the bovine serum albumin is 20 g / L; In step ⑤, in the mixed solution IV, the concentration of the buffer V is 30 mM, the concentration of the trehalose is 35 g / L, and the concentration of the glycerol is 35 g / L.

10. A coronavirus rapid detection device, characterized in that: The coronavirus rapid detection device is provided with a light source, and is characterized in that the coronavirus rapid detection device is also provided with a slit, a grating, a detection mechanism, and a spectrometer. The detection mechanism is provided with a sample bottle for containing a liquid reagent to be tested; the light source outputs a stable light beam of a fixed wavelength, and the stable light beam enters the grating after passing through the slit, and after being split by the grating, it passes through the sample bottle and the liquid reagent to be tested inside it to form a transmitted light beam, and the transmitted light beam enters the spectrometer to detect the light intensity; the width of the slit is adjustable, and the adjustment range is 0 to 5 mm; the detection mechanism is also provided with a platform base, and the platform base is used to place the sample bottle.

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