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Dye liquid special for preparation of high-resolution chromosome by karyotype analysis

A karyotype analysis, high-resolution technology, applied in the biological field, can solve the problems of unsatisfactory effect, affecting the appearance of banding, poor dyeing texture effect, etc., to improve uniformity and clarity, improve dispersion characteristics, and clear dyeing effect. Effect

Active Publication Date: 2019-03-29
BEIJING INST OF HUMAN GENETICS & REPRODUCTION MEDICINE LTD CHINA +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the Giemsa reagents of various manufacturers have unsatisfactory results when doing high-resolution and super-high-resolution karyotypes. The performance is: long staining time, uneven banding, and poor staining texture.
Because the preparation of the existing G banding dye solution is not standardized enough, the banding is blurred, which affects the appearance of the banding, such as patents CN 103710435 B and CN 103308361 B
This brings difficulties to the analysis of chromosome karyotype results, affecting the quality of chromosome preparation and the external quality assessment of karyotype analysis results

Method used

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  • Dye liquid special for preparation of high-resolution chromosome by karyotype analysis
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  • Dye liquid special for preparation of high-resolution chromosome by karyotype analysis

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Effect test

Embodiment 1

[0043] The preparation method of the special high-resolution chromosome dye solution for karyotype analysis of the present invention:

[0044] 1. Dye solution formula

[0045] Giemsa powder 1g, glycerin 66ml, methanol 66ml.

[0046] 2. Preparation method

[0047] (1) Mix 1 g of Giemsa powder and 6 ml of glycerin, and grind continuously for 2 hours in the dark.

[0048] (2) Transfer the Giemsa grinding mixture into a 100ml brown bottle, and add 60ml of glycerin. Then incubate at 58°C in the dark for 2 hours.

[0049] (3) Then transfer to a 500ml brown bottle, add 66ml methanol and mix well. After being sealed and protected from light and stored at room temperature for 2 weeks, the performance test of Giemsa dye solution can be carried out.

[0050] (4) Dye liquor performance test: use G-belt dyeing technology to test the dye liquor performance:

[0051] A. Cylinder No. 1: Put the baked human peripheral blood sample slides into the trypsin mixture and digest at 37°C;

[0...

Embodiment 2

[0068] The preparation method of the special high-resolution chromosome dye solution for karyotype analysis of the present invention:

[0069] 1. Dye solution formula

[0070] Giemsa powder 1g, glycerin 66ml, methanol 66ml.

[0071] 2. Preparation method

[0072] (1) Mix 1 g of Giemsa powder and 6 ml of glycerin, and grind continuously for 2 hours in the dark.

[0073] (2) Testing indicators: After grinding, the next step will start only after the particle size of the Giemsa dyeing powder has been tested twice and passed. Specific steps are as follows:

[0074] A. Manual detection step 1: Take 10 μl of Giemsa grinding mixture and apply it on the glove and twist it. If the particles are visible to the naked eye, and if the particles are touched by finger twisting, the grinding is not sufficient. Continue grinding in step (1) until it cannot be touched. until particles are present.

[0075] B. Microscopic detection step 2: After no particles can be found by twirling, under ...

Embodiment 3

[0088] The preparation method of the special high-resolution chromosome dye solution for karyotype analysis of the present invention:

[0089] 1. Dye solution formula

[0090] Giemsa powder 1g, glycerin 66ml, methanol 66ml, dispersant.

[0091] Wherein, described dispersant:

[0092] The dispersant of experimental group 1 is ethylene oxide with a working volume of 1ml.

[0093] The dispersant of experimental group 2 is ethylene oxide with a working volume of 2.5ml.

[0094] The dispersant of experimental group 3 is ethylene oxide with a working volume of 5ml.

[0095] The dispersant of experimental group 4 is ethylene oxide with a working volume of 10ml.

[0096] The dispersant of comparative group 1 is ethylene oxide with a working volume of 0ml.

[0097] 2. Preparation method

[0098] (1) Mix 1 g of Giemsa powder and 6 ml of glycerin, and grind continuously for 2 hours in the dark.

[0099] (2) Testing indicators: After grinding, the next step will start only after th...

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Abstract

The invention relates to a dye liquid special for preparation of high-resolution chromosome by karyotype analysis. The dye liquid is prepared by the following raw materials according to a proportion:1-3g of Giemsa dye power, 66-69ml of glycerinum, 66ml of methyl alcohol and 1-10 ml of dispersing agent. The dye particle is more refined and standard by grinding the dye particle, the dispersing characteristic of the dye particle is improved by addition of a dispersing agent, so that the uniformity and the clearness of dyeing are improved. By the novel dye liquid, the banding dyeing quality of the high-resolution chromosome is obviously improved, the banding belt dyeing effect of the high-resolution chromosome is clearer, the dyeing speed of human chromosome is rapid and controllable, and thebelt is clear. The dye liquid can be applied to large-scale clinic genetic laboratory.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the special preparation of high-resolution chromosome dye solution for karyotype analysis. Background technique [0002] Chromosomes are the carriers of genes. Chromosomes may undergo quantitative and structural changes, which may cause various genetic defects, congenital genetic diseases, embryo cessation, spontaneous abortion, tumors, etc. Diseases caused by chromosomal abnormalities are generally called chromosomal diseases. [0003] Chromosomal detection is currently mostly used in clinical practice such as reproduction, obstetrics and gynecology, newborns, blood diseases, tumors and other specialties. Genetic diagnosis using the technique of chromosome banding after cell culture is relatively simple and reliable, and it is still the gold standard test method. [0004] Chromosomal karyotype analysis is mainly divided into several steps: cell culture, harvest, preparation, bandi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
CPCG01N1/30
Inventor 李存玺刘恒涛刘舒李能干
Owner BEIJING INST OF HUMAN GENETICS & REPRODUCTION MEDICINE LTD CHINA
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