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Joint detection kit, detection method and immunoassay system

A combined detection and kit technology, applied in the field of immune detection, can solve the problems of consumption, increased cost, hospital and patient burden, and achieve the effect of improving detection efficiency

Inactive Publication Date: 2021-06-29
SHENZHEN DYMIND BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] There are currently many ways to detect PCT and IL-6, but it is impossible to realize the joint detection of the two items in a single channel. If you want to refer to the results of PCT and IL-6 two inflammatory indicators, you need to consume more samples. and detection time, which increases the cost and brings greater burden to the hospital and patients

Method used

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  • Joint detection kit, detection method and immunoassay system
  • Joint detection kit, detection method and immunoassay system

Examples

Experimental program
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Effect test

Embodiment 1

[0076] Preparation of capture antibody composition

[0077] 1) Select two types of microspheres with a particle size of 5.5 μm and 8 μm, the 5.5 μm microspheres are coated with mouse anti-human PCT antibody, and the 8 μm microspheres are coated with mouse anti-human IL-6 antibody;

[0078] 2) 1 mg of magnetic fluorescent-encoded microspheres were magnetically separated using a magnetic separation plate, and the supernatant was absorbed;

[0079] 3) Add 1 mL of 50 mM, pH 6.0 2-morpholineethanesulfonic acid (MES) buffer solution, vortex the microspheres for 20 s, perform magnetic separation with a magnetic separation plate, and absorb the supernatant. This step is repeated three times;

[0080] 4) Add 2 mL of the above-mentioned MES buffer solution, vortex the microspheres for 20 seconds, and add 10 μL of 50 mg / mL N-hydroxysuccinimide (NHS) and 1-(3- Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) solution, incubate with shaking at room temperature for 20 min, per...

Embodiment 2

[0096] Preparation of capture antibody composition

[0097] 1) Two kinds of encoded magnetic fluorescent microspheres with different encodings are selected and coated with mouse anti-human PCT antibody and mouse anti-human IL-6 antibody respectively;

[0098] 2) 0.5 mg of magnetic fluorescence-encoded microspheres were magnetically separated using a magnetic separation plate, and the supernatant was absorbed;

[0099] 3) Add 1mL of 20mM, pH 7.0 N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid (HEPES) buffer solution, vortex the microspheres for 20s, and use a magnetic separation plate Magnetically separate and aspirate the supernatant. This step is repeated three times;

[0100]4) Add 1 mL of the above-mentioned HEPES buffer, vortex the microspheres for 20 seconds, add 5 μL of 50 mg / mL N-hydroxysuccinimide (NHS) and 1-(3 -Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) solution, then add 20 μg of the corresponding antibody to the solution, incubate with sh...

Embodiment 3

[0115] Preparation of capture antibody composition

[0116] 1) Two kinds of encoded magnetic fluorescent microspheres with different encodings are selected and coated with goat anti-human PCT antibody and rabbit anti-human IL-6 antibody respectively;

[0117] 2) 1.0 mg of magnetic fluorescence-encoded microspheres were magnetically separated using a magnetic separation plate, and the supernatant was absorbed;

[0118] 3) Add 1 mL of 20 mM boric acid (BS) buffer solution with pH 8.0, vortex the microspheres for 20 s, perform magnetic separation with a magnetic separation plate, and absorb the supernatant. This step is repeated three times;

[0119] 4) Add 2 mL of the above-mentioned boric acid buffer solution, vortex the microspheres for 20 seconds, add 5 μL of 100 mg / mL N-hydroxysuccinimide (NHS) and 1-(3 -Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) solution, then add 20 μg of the corresponding antibody to the solution, incubate with shaking at room temperat...

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Abstract

The invention discloses a joint detection kit, a detection method and an immunoassay system.The joint detection kit comprises a capture antibody composition, a detection antibody composition and a marker. The capture antibody composition comprises a first solid-phase carrier, a first capture antibody connected with the first solid-phase carrier, a second solid-phase carrier, and a second capture antibody connected with the second solid-phase carrier. The detection antibody composition comprises a first detection antibody and a second detection antibody; the marker comprises a first marker and a second marker; and the first solid-phase carrier is different from the second solid-phase carrier, and is used for distinguishing a compound formed by connecting with the first solid-phase carrier and a compound formed by connecting with the second solid-phase carrier during detection. Through the mode, the procalcitonin and the interleukin 6 in a to-be-detected sample can be detected at the same time so that the detection efficiency is improved.

Description

technical field [0001] The present application relates to the technical field of immunoassay, in particular to a combined detection kit, a detection method, and an immunoassay system. Background technique [0002] There is only a small amount of procalcitonin (PCT) in the blood of healthy people, and PCT will increase significantly after bacterial infection, and it is mainly used for auxiliary diagnosis of bacterial infectious diseases clinically; Interleukin-6 (Interleukin-6, IL-6) is a pleiotropic cytokine with many functions. After a single endotoxin stimulation, IL-6 is the earliest marker to increase, and it can play an early warning role for bacterial infection. It is mainly used clinically to monitor the body's immune status, inflammatory response, etc. [0003] There are currently many ways to detect PCT and IL-6, but it is impossible to realize the joint detection of the two items in a single channel. If you want to refer to the results of PCT and IL-6 two inflamma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74G01N33/68G01N33/543
CPCG01N33/74G01N33/6869G01N33/543G01N2333/585G01N2333/5412
Inventor 许墨横马志亚李春晖陆锋
Owner SHENZHEN DYMIND BIOTECH