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Manufacturing method of peripheral blood chromosome G band

A production method and chromosome technology, applied in the field of medical examination, can solve the problems of lowering the success rate of chromosome karyotype analysis, lowering clinical practicability, small size of analysable split images, etc., achieving reliable diagnosis results, lowering production costs, and good dispersion. Effect

Pending Publication Date: 2021-09-28
北京明基乐威生物科技有限公司
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AI Technical Summary

Problems solved by technology

Due to the complex process of making peripheral blood chromosomes and many influencing factors, there are often phenomena such as small analyzable mitotic figures, thick and short chromosomes, and poor dispersion, which greatly reduces the success rate of chromosome karyotype analysis and reduces its clinical practicability. Routine chromosome analysis is difficult to detect abnormalities < 10M bases

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  • Manufacturing method of peripheral blood chromosome G band
  • Manufacturing method of peripheral blood chromosome G band
  • Manufacturing method of peripheral blood chromosome G band

Examples

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Embodiment 1

[0038] Embodiment 1: Production of Peripheral Blood Chromosome G Band The specific production method includes the following steps:

[0039] (1) Preparation of culture medium: Take out the peripheral blood culture medium from the freezer, incubate in a 37°C water bath, and set aside.

[0040] (2) Inoculation: the inoculation amount of peripheral blood per 5 ml of peripheral blood cell culture medium is 0.5 ml.

[0041] (3) Cultivation: Cultivate in an incubator at 37° C. for 66-72 hours.

[0042] (4) Termination of culture: Before harvesting, add 100 μl of 10 μg / ml colchicine to each bottle, and then put them into an incubator at 37°C to stop cell division and make them stay in the metaphase of cells, and harvest after 3 hours.

[0043] (5) Collection of peripheral blood cell culture:

[0044] a. Centrifuge: 1500rpm / min, 5 minutes, discard the supernatant;

[0045] b. Hypotonicity: Add about 5ml of 0.075M KCL hypotonic solution pre-warmed to 37°C, mix gently, and place at 34...

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Abstract

The invention relates to a manufacturing method of a high-resolution peripheral blood chromosome G band. The manufacturing method comprises the following steps: (1) inoculating peripheral blood into a peripheral blood cell culture medium and culturing; (2) adding colchicine into the peripheral blood cell culture medium, and terminating culture; 0.4 ug / ml colchicine is added into each 5m of the peripheral blood cell culture medium; (3) collecting a peripheral blood cell suspension, and adjusting the concentration of the cell suspension to 1.2-3.6 * 10 < 9 > / L; (4) dripping the cell suspension on a slide, and drying in an oven at 75 DEG C for 3 hours; (5) carrying out zoning treatment on the dried slide for 6-10 minutes by using trypsin with the concentration of 0.01%; and (6) dyeing for 4 minutes and 30 seconds by adopting a Giemsa dye to obtain a chromosome specimen with G banding. The peripheral blood chromosome manufactured by the method is in a long rod shape, is well dispersed without crossing, has clear banding and is high in banding level.

Description

technical field [0001] The invention relates to the technical field of medical testing, in particular to a method for making a peripheral blood chromosome G-band. Background technique [0002] Chromosomes are the carriers of genes, and chromosomal abnormalities will inevitably cause unbalanced gene expression, thereby causing a series of clinical symptoms. Studies have shown that chromosomal abnormalities are closely related to blood system diseases, mental retardation, and reproductive abnormalities. In the field of human reproductive research, diseases such as miscarriage, stillbirth, fetal arrest, infertility, azoospermia, premature ovarian failure, immature uterus, birth defects, and hypoplasia of secondary sexual characteristics are related to structural abnormalities, abnormal numbers, and polymorphisms of chromosomes. Therefore, it is necessary to perform karyotype analysis on such patients. In 1968, the Swedish cytochemist Caspersson et al. applied the fluorescent ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/31G01N1/40G01N1/44
CPCG01N1/31G01N1/4077G01N1/44G01N2001/4083
Inventor 不公告发明人
Owner 北京明基乐威生物科技有限公司
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