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Rapid detection and diagnosis method for biological activity of in-vitro trace follicle-stimulating hormone

A biological activity, follicle-stimulating hormone technology, applied in the field of rapid detection and diagnosis of the biological activity of trace follicle-stimulating hormones in vitro, can solve the problem of no good feasible method and the like, and achieve the effect of good durability

Pending Publication Date: 2022-03-11
NANTONG UNIVERSITY
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Problems solved by technology

[0005] The purpose of the present invention is to solve the problem of in vitro diagnosis in the prior art, especially when the amount of FSH obtained from the patient is very limited, and there is no good feasible method for the detection and diagnosis of its biological activity. question

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  • Rapid detection and diagnosis method for biological activity of in-vitro trace follicle-stimulating hormone
  • Rapid detection and diagnosis method for biological activity of in-vitro trace follicle-stimulating hormone
  • Rapid detection and diagnosis method for biological activity of in-vitro trace follicle-stimulating hormone

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Embodiment Construction

[0031] In order to make the objects, technical solutions, and advantages of the present invention, the present invention will be further described in detail below with reference to the specific embodiments.

[0032] Many specific details are set forth in the following description to fully understand the present invention, but the present invention can also be implemented in other than described in this manner, and therefore, the present invention is not limited to the specific embodiments of the specification below. limit.

[0033] See figure 1 , A rapid detection and diagnostic method of biological activity in vitro, including the following steps:

[0034] S1: Sampling: Concentrate from a small amount of urine samples FSH;

[0035] Specifically, in one embodiment, 30 ml of urine is acquired from the object to be examined, and the filter paper is simplely filtered to remove the cell tissue fragment;

[0036] The urine is then transferred to two 15 ml ultrafiltration tubes, and the...

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Abstract

The invention provides a method for rapidly detecting and diagnosing biological activity of in-vitro trace follicle stimulating hormone, which relates to the technical field of biomedicine and comprises the following steps: concentrating FSH (follicle stimulating hormone) from a small amount of urine sample; further removing LH from the concentrated FSH sample to obtain purified FSH; the method comprises the following steps: establishing an SNAP-FSHR HEK 293 stable expression cell strain; stimulating FSHR internalization by using the sample FSH; stimulating the FSHR by using the standardized FSH and causing in vivo retrogression of the receptor; acquiring an FSHR internalized image by utilizing microphotography; and calculating the number of internalized receptor particles by using Image J high-content image analysis software, and comparing the biological activity of the sample to be detected and the biological activity of the standard substance. The invention establishes and verifies a method for rapidly determining the biological activity of trace FSH in vitro by further analyzing high-content images by utilizing the reaction of FSHR stably expressed by a bioengineering cell strain to FSH. The FSH with LH removed can be rapidly recovered from human urine, and the recovery rate is about 90%. The internalization of the FSHR expressed in the HEK293 cell has a very good dose response relationship to trace FSH. The linear range is 0.01-50 ng / mL, and the correlation coefficient R2 is greater than or equal to 0.99.

Description

Technical field [0001] The present invention relates to the field of biomedical technology, and more particularly to a rapid detection and diagnosis method of biological activity in vitro microcanodoxin. Background technique [0002] Macrocarcin (FSH) has a very important role in maintaining normal physiological and reproductive function, including promoting follicular growth, development, and matures with mysteria cells. FSH is synthesized by pituitary front leaf bacteric cells, secreted a type of glycoprotein, composed of α, β, α having 115 amino acids, β having a specificity of β subunits, molecular weight 30kd. [0003] Macrobin receptor (FSHR) is a G protein coupled receptor. FSH is activated by activating FSHR, promoting follicular peripheral cells into internal and outer laminate cells and synthesized to proximal and yellow biochemical receptors through downstream signaling pathways; Transforming androgen into a female, and promoting the secretion of follicles and increase...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/493G01N1/34G01N1/40
CPCG01N33/493G01N1/34G01N1/40
Inventor 刘建喜
Owner NANTONG UNIVERSITY
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