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An ELISA detection kit for human asprosin protein and its use

A detection kit and human-derived technology, applied in the biological field, can solve the problem of no clear diagnosis of PCOS patients, and achieve the effects of high practical value, high sensitivity, and easy popularization and application.

Inactive Publication Date: 2018-09-14
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

[0012] The original intention of the present invention is to overcome the problem that there is still no definite diagnosis of PCOS patients at present, and propose a human-derived asprosin protein ELISA detection kit and its application

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  • An ELISA detection kit for human asprosin protein and its use
  • An ELISA detection kit for human asprosin protein and its use
  • An ELISA detection kit for human asprosin protein and its use

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Embodiment Construction

[0056] According to the gene sequence of protein A and the expression preference of E. coli prokaryotic protein, on the premise of ensuring the consistent amino acid sequence of gene A, optimize the original codon of gene A, and design primers, original codon, optimized codon, See Table 1 for primers;

[0057] Table 1. Asprosin protein gene and amino acid sequence

[0058]

[0059] figure 1 It is SDS-PAGE analysis of protein A after induction under different conditions, where M represents protein marker; Lane 1 is the total protein expressed before induction; Lane 2 is the protein in the supernatant after the cells were broken after induction at 20°C overnight; lane 3 is the protein in the precipitate after induction at 20°C overnight; lane 4 is the protein in the supernatant after induction at 37°C for 4 hours; lane 5 is the precipitation after induction at 37°C for 4 hours protein in.

[0060] figure 2 SDS-PAGE analysis of asprosin protein after purification by nick...

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Abstract

The invention provides an ELISA (enzyme-linked-immunosorbent serologic assay) detection kit of a human asprosin protein and a use of the kit. The kit comprises the following components: an ELISA plate, a standard product, a negative reference product, a positive reference product, a sample dilution solution, a washing buffer solution, a second antibody, an antibody dilution solution, developing liquid and terminating liquid; the ELISA plate is enveloped by a specific polyclonal antibody of the human asprosin protein with a biological element label and closed by virtue of a closed buffer solution; and the second antibody can be specifically combined with the polyclonal antibody as a first antibody. The kit can rapidly detect the content of the asprosin protein in a human or an animal specimen, and is less in time consumption, simple and convenient in use, high in result specificity and high in sensitivity. On the basis, the invention also establishes a detection device system for assisting the diagnosis of PCOS (polycystic ovary syndrome), and the detection device system only needs to be equipped with a 37-DEG C incubator, a pipettor and a microplate reader.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to an ELISA detection kit for human asprosin protein. Background technique [0002] Polycystic ovary syndrome (PCOS) is the most common reproductive disorder and metabolic syndrome. It has become one of the research hotspots and difficulties in the field of gynecological endocrinology. At present, the etiology and pathogenesis of PCOS are still unclear. Insulin resistance (IR) and hyperandrogenemia (hyperandrogenemia, HA) are the main pathological features of PCOS, which are often accompanied by obesity, abnormal glucose metabolism, and cardiovascular diseases. occurrence of the disease. Due to the heterogeneity of PCOS and the polymorphism of clinical manifestations, it brings great difficulties to clinical diagnosis and treatment. The diagnosis of PCOS has always been a very controversial topic. [0003] At present, three PCOS diagnostic criteria have been widely used in the world....

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/543G01N33/532
CPCG01N33/532G01N33/543G01N33/6893
Inventor 李博陈书强王晶晶
Owner FOURTH MILITARY MEDICAL UNIVERSITY