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Homogeneous joint detection reagent and method based on immunoturbidimetry and afterglow luminescence

A combined detection and reagent technology, applied in the field of immunodetection, can solve problems such as difficulty in completing accurate detection, and achieve the effects of reducing time, improving detection sensitivity, and good repeatability

Pending Publication Date: 2020-07-31
SHANGHAI TAYWELL BIOTECHNOLOGY CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is difficult to complete the accurate detection of these two items in a single sampling

Method used

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  • Homogeneous joint detection reagent and method based on immunoturbidimetry and afterglow luminescence
  • Homogeneous joint detection reagent and method based on immunoturbidimetry and afterglow luminescence
  • Homogeneous joint detection reagent and method based on immunoturbidimetry and afterglow luminescence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0233] Simultaneous detection of SAA and CRP in whole blood samples using the homogeneous no-wash combined detection method of the present invention

[0234] 1.1 Introduction of substances used

[0235] Samples to be tested: 50 random samples were taken at one time, including whole blood samples of normal people and whole blood samples of patients with inflammation;

[0236] Light absorbing agent: phthalocyanine dye, excitation wavelength 730nm;

[0237] Luminescent agent: europium complex, luminous wavelength 615nm;

[0238]

[0239] Photochemical buffer:

[0240]

[0241] Counterparts that specifically bind to biomarkers: SAA-Ab1 monoclonal antibody, SAA-Ab2 monoclonal antibody, CRP polyclonal antibody

[0242] Carrier Microspheres: Carboxylated Polystyrene Balls

[0243] 1.2, the preparation of turbidity reagent I

[0244] Dissolve 5 mL of carboxypolystyrene spheres with a particle diameter of 130 nm and a solid content of 5% in 100 mL of ultrapure water, and u...

Embodiment 2

[0300] Simultaneous detection of CRP and PCT in whole blood samples using the homogeneous wash-free combined detection method of the present invention

[0301] 2.1 Introduction of the substances used

[0302] Samples to be tested: 50 random samples were taken at one time, including whole blood samples of normal people and whole blood samples of patients with inflammation;

[0303] Light absorbing agent: phthalocyanine dyes, excitation 680nm;

[0304] Luminescent agent: BODIPY, luminescence wavelength 540nm;

[0305] Photochemical buffer:

[0306]

[0307] Counterparts that specifically bind to biomarkers: PCT-Ab1 monoclonal antibody, PCT-Ab2 monoclonal antibody, CRP polyclonal antibody

[0308] Carrier Microspheres: Aldehydated Polystyrene Balls

[0309] 2.2, the preparation of turbidity reagent I

[0310] Repeat the procedure of step 1.2 in Example 1 to prepare turbidity reagent I, wherein 5 ml of aldylated polystyrene balls with a particle diameter of 130 nm and a ...

Embodiment 3

[0354] Simultaneous detection of CRP and IL-6 in whole blood samples using the homogeneous no-wash combined detection method of the present invention

[0355] Together with PCT, IL-6 and CRP are one of the main inflammatory markers and also one of the main biomarkers of sepsis. In clinical application, IL-6 mainly cooperates with CRP and PCT to monitor the postoperative inflammation of patients, the effect of medication and the healing status of patients. In addition, IL-6 can also be used to judge the occurrence and damage degree of rheumatoid arthritis. The reference value of IL-6 is 50-150pg / ml, which requires extremely high sensitivity of the detection method. However, the joint detection of IL-6 and CRP can be realized by adopting the method according to the present invention, wherein the highly sensitive IL-6 concentration can be reflected by the afterglow luminescence signal value, while the CRP concentration can be represented by the turbidimetric signal value.

[...

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Abstract

The invention relates to a homogeneous joint detection reagent composition. The composition comprises a turbidity reagent I and a luminescent reagent II, wherein the turbidity reagent I comprises a carrier microsphere connected with a counterpart capable of being specifically binding to a turbidimetric biomarker, and the turbidity reagent I and the turbidimetric biomarker are subjected to an immune reaction to increase the turbidity; and the luminescent reagent II comprises a donor component II-1 and an acceptor component II-2, wherein the donor component II-1 comprises a donor microsphere connected with a counterpart specifically binding to an afterglow biomarker, the donor microsphere generates singlet oxygen under light excitation, and the acceptor component II-2 comprises an acceptor microsphere connected with a counterpart specifically binding to the afterglow biomarker, and the acceptor microsphere reacts with the singlet oxygen to generate an afterglow luminescence signal. In addition, the invention also relates to a homogeneous joint detection method and a kit containing the reagent composition.

Description

[0001] field of invention [0002] The invention relates to the technical field of immunoassay, in particular to a homogeneous combined detection reagent composition, kit and corresponding detection method based on immunoturbidimetry and afterglow luminescence. [0003] Background of the invention [0004] In vitro diagnostics are the main approach used today to diagnose various diseases. In addition to disease diagnosis, in vitro diagnostic reagents are also used for detection and early warning of personal health. The scope of in vitro diagnostic reagents is extremely wide, including a series of items such as liver and kidney function, myocardial infarction detection and inflammation diagnosis. In vitro diagnosis mostly uses blood, urine, and saliva as test samples in order to achieve rapid, convenient, and accurate diagnosis. [0005] In the field of in vitro diagnostics, immunoassay technology is an important component. Immunoassay technology is a detection method based o...

Claims

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Application Information

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IPC IPC(8): G01N33/539G01N33/533G01N33/543G01N21/64C09K11/06
CPCG01N33/539G01N33/533G01N33/54313G01N21/64C09K11/06C09K2211/182C09K2211/1092C09K2211/1044C09K2211/1055C09K2211/1011
Inventor 葛霄鹏李颖严志伟
Owner SHANGHAI TAYWELL BIOTECHNOLOGY CO LTD
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