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Chemiluminiscence immunoassay kit for detecting 14-3-3[eta] protein and application thereof

A chemiluminescence immunoassay and detection kit technology, applied in the field of immunoassays, can solve the problems of not finding a 14-3-3eta protein detection kit, cumbersome operation, low accuracy and so on.

Pending Publication Date: 2020-01-24
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Existing methods for detecting 14-3-3 protein are heterogeneous immunoassays, which are less sensitive, less accurate, and more complicated to operate
Moreover, no commercial 14-3-3eta protein detection kit has been found in the domestic and foreign markets
It has been reported that enzyme-linked immunosorbent assay (ELISA) was used to detect 14-3-3eta protein in serum of patients, but its sensitivity is low, specificity is poor, and the operation is relatively cumbersome.

Method used

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  • Chemiluminiscence immunoassay kit for detecting 14-3-3[eta] protein and application thereof
  • Chemiluminiscence immunoassay kit for detecting 14-3-3[eta] protein and application thereof
  • Chemiluminiscence immunoassay kit for detecting 14-3-3[eta] protein and application thereof

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Experimental program
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Effect test

Embodiment approach

[0112] As mentioned above, the existing methods for detecting 14-3-3 protein are all heterogeneous immunoassay methods, which have low sensitivity, low accuracy and complicated operation. The inventors found that the detection of 14-3-3 protein by homogeneous chemiluminescence immunoassay has high sensitivity, wide linear range and good precision, and the detection process does not require steps such as washing, and the operation is relatively simple. The present invention has been made based on the above findings.

[0113] Therefore, the present invention relates to a chemiluminescent immunoassay kit for detecting 14-3-3 eta protein, which comprises: a first reagent, a second reagent, a calibrator, a low-level quality control product and a high-level quality control product , wherein: the first reagent comprises luminescent particles coated with a first anti-14-3-3eta protein antibody, and the first anti-14-3-3eta protein antibody can bind to the first expression of 14-3-3eta...

specific Embodiment

[0307] In order to make the present invention easier to understand, the present invention will be further described in detail below in conjunction with examples, and these examples are for illustrative purposes only, and do not limit the scope of application of the present invention. The raw materials or components used in the present invention can be prepared by commercial channels or conventional methods unless otherwise specified.

[0308]In the method of the present invention, after all reagents are combined or mixed, they can be uniformly mixed and / or incubated according to actual needs. Specifically, the incubation temperature may be any temperature within the temperature range of 25-45° C., and the incubation time may be overnight or 10-20 minutes.

Embodiment 1

[0310] 1. Preparation of the first reagent (luminescent particles coated with anti-14-3-3eta protein antibody):

[0311] (1) Preparation of luminescent particles and photosensitive particles

[0312] For the preparation method, composition structure and content of the luminescent particles and photosensitive particles used in the present invention, please refer to Example 1 in Chinese Patent CN100429197C (this patent document is hereby incorporated by reference in its entirety).

[0313] (2) Pretreatment

[0314] Put 0.2 mg of the antibody raw material to be treated into a dialysis bag (molecular weight cut-off of 14KD), put the dialysis bag into a beaker, add 100 times the volume of 0.05M pH9.6 CB dialysis buffer into the beaker, and place it on a magnetic stirrer. Dialysis was performed at 2-8°C. Change the dialysate at least once, and dialyze for at least 4-5 hours each time. Aspirate the dialyzed protein and transfer it to a clean centrifuge tube, and take a sample to d...

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Abstract

The invention relates to a chemiluminiscence immunoassay kit for detecting 14-3-3[eta] protein. The kit is prepared from a first anti-14-3-3-[eta] protein antibody and a second anti-14-3-3-[eta] protein antibody which can be specifically combined with 14-3-3-[eta] protein; and moreover, by adopting a double-antibody sandwich mode and utilizing the advantages of the high sensitivity, the wide linear range, the good precision and the like of a light-activated chemiluminescence immunoassay platform, the definite diagnosis rate is improved for clinical diagnosis of rheumatoid arthritis, and auxiliary detection is provided for prevention of rheumatoid arthritis and early discovery of rheumatoid arthritis.

Description

technical field [0001] The invention belongs to the technical field of immune analysis, and in particular relates to a chemiluminescence immunoassay kit for detecting 14-3-3 eta protein, a preparation method and a use method thereof. Background technique [0002] 14-3-3eta(η) protein is a novel serum / plasma protein marker that induces inflammatory factors such as interleukin (IL)-1 and -6 and is linked to joint damage. Among the seven subtypes, only 14-3-3η protein was highly expressed in RA active inflammatory synovium and serum, and was positively correlated with the expression of RA pro-inflammatory factors and synovial inflammation, and the level of 14-3-3η protein in synovial fluid At least 5 times higher than the serum level, suggesting that synovium is the main source of 14-3-3η protein. 14-3-3η isoforms are expressed at high levels in arthritic patients compared to healthy individuals, which is thought to be directly related to the ability of 14-3-3η proteins to ind...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/6803G01N33/54313
Inventor 饶星廖智星刘宇卉李临其他发明人请求不公开姓名
Owner CHEMCLIN DIAGNOSTICS CO LTD
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