Diagnosis quality-control cell strain for common numerical abnormalities of chromosomes and preparation method thereof

A technology for chromosome number and abnormal diagnosis, applied in biochemical equipment and methods, cells modified by introducing foreign genetic material, using vectors to introduce foreign genetic material, etc., can solve the problem of no feasible quality control substances

Inactive Publication Date: 2014-09-24
翁炳焕
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Problems solved by technology

[0011] In order to solve the problem that there is no feasible quality control substance in prenatal diagnos

Method used

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  • Diagnosis quality-control cell strain for common numerical abnormalities of chromosomes and preparation method thereof
  • Diagnosis quality-control cell strain for common numerical abnormalities of chromosomes and preparation method thereof

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Embodiment Construction

[0019] 1. Primordial cells: The primitive cells used are derived from trisomy 21, trisomy 18, trisomy 13, X or Y chromosome after diagnosis of chromosomal disease due to the needs of clinical diagnosis and treatment and have been diagnosed as one of the common abnormalities in chromosome number Abnormal remaining primary or passage living tissue cells with adherent growth; the primary or passage living tissue cells may also involve living tissue cells with abnormal chromosome structure or No. 1 to No. Live tissue cells with abnormal number of chromosome 22. The cell types are fetal amniotic fluid, villous tissue and other cells.

[0020] 2. Extraction of SV40 large T antigen DNA: ① SV40 DNA digestion: buy SV40 freeze-dried powder or SV40 plasmid containing large T antigen gene from the market, dissolve in appropriate amount of H 2 In O or TE buffer, add 2uL 10× digestion buffer and 18uL H 2O, add restriction endonuclease BamH I (1-5U / ugDNA), incubate at 37°C for 1h, heat at ...

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Abstract

The invention relates to a diagnosis quality-control cell strain for common numerical abnormalities of chromosomes and a preparation method thereof. The invention is mainly characterized in that the preparation method comprises: taking residual attached living cells which are used for chromosomal disease diagnosis as needed by clinical diagnosis and treatment and are definitely diagnosed to have 21-trisomy, 18-trisomy, 13-trisomy, X or Y chromosome abnormalities, introducing, through transgenosis, SV40LTag-pcDNA3.1(-) plasmid constructed by SV40LTag DNA and pcDNA3.1(-) connected by T4DNA ligase and digested by BamHI, allowing the plasmid to integrate with the cell DNA, screening cell strains integrated with recons by G418, performing subculture amplification and cryopreservation, then extracting the cell strains with numerical abnormalities of chromosomes, mixing the cell strains according to a required ratio for quality control so as to convert each original cell strain with a single numerical abnormality of chromosomes into a chimera quality-control cell strain with a certain ratio of five numerical abnormalities of chromosomes. Therefore, the difficulty for differential diagnosis and chimera diagnosis is increased; the cell strain is used as a quality-control cell strain for diagnosis of common numerical abnormalities of chromosomes; samples are easily available; and waste residual cells are converted into effective immortalized quality-control cells.

Description

technical field [0001] The invention relates to a quality control cell line for the diagnosis of common chromosome number abnormalities and a preparation method thereof, which are mainly used in prenatal diagnosis laboratories in the medical field for technical assessment, indoor quality control and external quality assessment of common chromosome number abnormalities diagnosis. Background technique [0002] Chromosomes are the genetic material in the nucleus. Humans have 23 pairs of chromosomes, 22 of which are autosomes, and 1 pair is the sex chromosome that determines the sex of men and women. Genes carrying genetic information on chromosomes, of which DNA accounts for more than 90%, and RNA content varies with cell cycle and growth, generally accounting for 1% to 10%. [0003] Chromosomal structural abnormalities, abnormal numbers, and mosaicism are clinically manifested as chromosomal diseases, which are common birth defects, such as congenital stupidity, primary amenor...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N15/85C12Q1/68
Inventor 翁炳焕
Owner 翁炳焕
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