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Homogeneous chemiluminescence POCT detection method and device using same

A homogeneous chemiluminescence, detection method technology, applied in chemiluminescence/bioluminescence, measurement devices, analysis by chemical reaction of materials, etc., can solve the problems of poor detection precision, low detection sensitivity, insufficient reaction, etc. , to achieve the effect of improving accuracy, high sensitivity and reducing non-specific adsorption

Pending Publication Date: 2020-01-31
BEYOND DIAGNOSTICS (SHANGHAI) CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But because these two kinds of technologies mainly carry out release detection on NC film, because the CV of film itself just has more than 5%, so the POCT detection CV of solid-phase film method generally all will be more than 10%, and detection precision is very poor. , for items with high sensitivity requirements such as cTnI, quantification becomes extremely difficult
In addition, the use of new technologies such as microfluidic chip type POCT detection technology has the advantages of fast response speed and small sample demand, but it also has the problem of low detection sensitivity due to insufficient reaction.

Method used

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  • Homogeneous chemiluminescence POCT detection method and device using same
  • Homogeneous chemiluminescence POCT detection method and device using same
  • Homogeneous chemiluminescence POCT detection method and device using same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0187] Example 1: Preparation of acceptor microspheres

[0188] 1. Prepare a 25mL round-bottomed flask, add 0.1g of europium(Ⅲ) complex, 10mL of 95% ethanol, stir magnetically, and raise the temperature of the water bath to 70°C to obtain a solution of europium(Ⅲ) complex.

[0189] 2. Prepare a 100mL three-neck flask, add 10mL 95% ethanol, 10mL water and 10mL polystyrene microspheres coated with carboxydextran hydrogel with a particle size of 200nm at a concentration of 10%, stir magnetically, and heat the water bath to 70 ℃.

[0190] 3. Slowly drop the europium(III) complex solution in step 1 into the three-necked flask in step 2, react at 70°C for 2 hours, stop stirring, and cool naturally to obtain an emulsion.

[0191] 4. Centrifuge the above emulsion for 1 hour at 30,000 g, discard the supernatant after centrifugation, and then resuspend with 50% ethanol. After repeated centrifugal washing for 3 times, it was resuspended with 50 mM CB buffer solution with a pH value of ...

Embodiment 2

[0193] Example 2: Receptor microspheres coated with antibodies

[0194] 1. Measure 10 mg of receptor microspheres coated with carboxydextran hydrogel in a centrifuge tube according to the prepared amount, and centrifuge at 10,000 rpm for 60 min.

[0195] 2. Discard the supernatant, add 2 mg of Anti-PCT antibody I to the precipitate (it can be the corresponding antibody embodiment of any other analysis item (anti-cTnI antibody I and anti-PCT antibody I)), 50 μL of Tween-20 ( 50mg / mL), and a certain volume of 0.05M MES pH=6.0 was added to make the final concentration of acceptor microspheres 10mg / mL.

[0196] 3. Ultrasound to mix quickly.

[0197] 4. Add 50 μL of NaBH to the centrifuge tube 3 CN (50mg / mL, prepared in 0.05M MES pH=6.0) was mixed evenly, and placed in a rotary mixer at 37°C for 36-48h.

[0198] 5. Blocking: add 1 mL of BSA (50 mg / mL, prepared in 0.05M MES pH=6.0), and place in a rotary mixer at 37°C for 12-16 hours.

[0199] 6. Washing: wash 3 times with 0.05M...

Embodiment 3

[0201] Example 3: Preparation of Donor Microspheres

[0202] 1. Prepare a 25mL round-bottomed flask, add 0.1g of copper phthalocyanine (II) and 10mL of DMF, stir magnetically, and raise the temperature of the water bath to 70°C to obtain a copper (II) phthalocyanine solution.

[0203] 2. Prepare a 100mL three-neck flask, add 10mL 95% ethanol, 10mL water and 10mL polystyrene microspheres coated with aldehyde dextran hydrogel with a particle size of 200nm at a concentration of 10%, stir magnetically, and heat the water bath to 70°C.

[0204] 3. Slowly drop the copper phthalocyanine (II) solution in step 1 into the three-necked flask in step 2, react at 70°C for 2 hours, stop stirring, and cool naturally to obtain an emulsion.

[0205] 4. Centrifuge the above emulsion for 1 hour at 30,000 g, discard the supernatant after centrifugation, and resuspend with 50% ethanol. After repeated centrifugal washing for 3 times, it was resuspended with 50 mM CB buffer solution with a pH valu...

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Abstract

The invention relates to a homogeneous chemiluminescence POCT detection method and a device using same. The method comprises the following steps: S1, mixing a to-be-detected sample with a receptor reagent and a donor reagent to form a to-be-detected mixture; S2, exciting the to-be-detected mixture to perform chemiluminescence by utilizing energy or an active compound, and immediately measuring thesignal intensity of chemiluminescence, wherein the donor reagent comprises donor microspheres, and the donor microspheres can generate active oxygen in an excited state; the receptor reagent comprises receptor microspheres, and the receptor microspheres can react with the active oxygen to generate a detectable chemiluminescence signal; and the particle size of the donor microspheres is greater than that of the receptor microspheres. The method has high sensitivity, high precision and wide range of a chemiluminescence analysis technology, and has the characteristics of rapidness, portability and the like of a POCT detection technology.

Description

technical field [0001] The invention belongs to the technical field of homogeneous chemiluminescence, and in particular relates to a homogeneous immunoassay POCT detection method and a system using the detection method. Background technique [0002] Homogeneous chemiluminescence analysis refers to a method for chemiluminescence detection without separating the complexes formed after binding and the remaining free reactants. [0003] The existing homogeneous chemiluminescence analysis has the following disadvantages: [0004] A. The instrument system is bulky and occupies a large area. At the same time, due to the large test throughput, the reagent card adopts 100 tests as the whole unit, which requires the laboratory sample size; [0005] B. The instrument system and reagents are expensive and the maintenance cost is high, so it is not suitable for primary medical institutions; [0006] C. The instrument is bulky and cannot be carried into the diagnosis and treatment site;...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/543
CPCG01N21/76G01N33/54346Y02A50/30
Inventor 杨阳赵卫国刘宇卉李临
Owner BEYOND DIAGNOSTICS (SHANGHAI) CO LTD
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