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Detection method of embryo secretory protein

A secretory protein and detection method technology, which is applied in the detection field of embryonic secretory protein, can solve the problem of low detection sensitivity, achieve high detection sensitivity, realize signal amplification, and have low versatility

Inactive Publication Date: 2020-03-17
深圳中山泌尿外科医院
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In order to solve the technical problem of low current detection sensitivity in the above-mentioned background technology, the present invention provides a detection method for embryo secreted protein

Method used

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  • Detection method of embryo secretory protein
  • Detection method of embryo secretory protein
  • Detection method of embryo secretory protein

Examples

Experimental program
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Effect test

Embodiment 1

[0049] A method for detecting embryo secreted protein, comprising the following steps:

[0050] a. Take the capture antibody and labeled antibody whose chorionic gonadotropin concentration of the embryo is o, connect the capture antibody to fluorescent magnetic beads, and label the labeled antibody with biotin. The excitation and emission wavelengths of the fluorescent magnetic beads are respectively 660nm and 690nm;

[0051] b. Mix the fluorescent magnetic beads in step a with the embryo culture solution with a secreted protein concentration of o and incubate at 37°C for 30 minutes, then take out the fluorescent magnetic beads and wash them for 3 to 5 times. After washing, the fluorescent magnetic beads and the labeled antibody Mix with streptavidin-labeled β-galactosidase and incubate at 37°C for 30 minutes, then take out the fluorescent magnetic beads and wash them 5 times, and resuspend the fluorescent magnetic beads with 10uL of PBS buffer after washing;

[0052] c. Put ...

Embodiment 2

[0061] The difference from Example 1 is that the concentration of chorionic gonadotropin in the embryo culture solution is 0.1 pg / mL.

[0062] The n is 137, the N is 9978, and the AEB is 0.0138 during the measurement of the multicolor laser-induced fluorescence detector.

Embodiment 3

[0064] The difference from Example 1 is that the concentration of chorionic gonadotropin in the embryo culture solution is 1 pg / mL.

[0065] During the measurement with a multicolor laser-induced fluorescence detector, n was 1135, N was 9871, and AEB was 0.1221.

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Abstract

The invention belongs to the field of a detection method and particularly relates to a detection method of embryo secretory protein. The detection method comprises steps of taking a capture antibody and a labeled antibody of to-be-detected embryo secretory protein, connecting the capture antibody with fluorescent magnetic beads, and labeling the antibody with biotin; then mixing the fluorescent magnetic beads with the to-be-detected embryo secretory protein and incubating; mixing the microdroplets with the labeled antibody and joint avidin labeled beta-galactosidase, performing incubating to form a magnetic bead immune complex, wrapping the magnetic bead immune complex and a reaction substrate in the microdroplets, performing incubating, and detecting fluorescence intensity by using a multi-color laser-induced fluorescence detector to obtain the content of the to-be-detected embryo secretory protein. The detection method is advantaged in that the enzymatic reaction is adopted to continuously catalyze substrate reaction to generate the fluorescent substance, signal amplification of the to-be-detected protein can be achieved, high detection sensitivity is achieved, and a technical problem of low detection sensitivity at present is solved.

Description

technical field [0001] The invention belongs to the field of detection methods, and in particular relates to a detection method of embryo secreted protein. Background technique [0002] Quantitative detection of a trace protein in complex biological samples is of great significance for the screening of clinical biomarkers, the monitoring of disease development process and the determination of disease development stage. A variety of proteins are secreted during embryonic growth and development, and these proteins are closely related to various functions during embryonic development and implantation. Microfluidics integrates the basic operation units such as sample preparation, reaction, separation, and detection in the biological, chemical, and medical analysis processes into a micron-scale chip to automatically complete the entire analysis process. Due to its great potential in the fields of biology, chemistry, and medicine, it has developed into a new research field interd...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N33/68G01N33/76
CPCG01N21/6402G01N21/6428G01N21/6486G01N33/689G01N33/76
Inventor 曾勇陈培林孙青熊风钟惠娴李观贵姚志鸿
Owner 深圳中山泌尿外科医院
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