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EBV transfected chimera quality-control cell for birth defect antenatal diagnosis and preparation method thereof

A prenatal diagnosis, B lymphocyte technology, applied in biochemical equipment and methods, cells modified by introducing foreign genetic material, and microbial assay/inspection, etc. To achieve the effect of improving the level of diagnosis and establishing a simple system

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

[0011] In order to solve the problem that the cytogenetic laboratory in the medical field does not have quality control materials for the technical assessment of common abnormal chromosome number diagnosis, indoor quality control and external quality assessment, the inventors proposed the present invention

Method used

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  • EBV transfected chimera quality-control cell for birth defect antenatal diagnosis and preparation method thereof

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Embodiment Construction

[0016] 1. Primordial cells: The primitive cells used are derived from trisomy 21, trisomy 18, trisomy 13, X or Y chromosomes after diagnosis of chromosomal diseases due to the needs of clinical diagnosis and treatment and have been diagnosed as one of the common abnormal chromosome numbers Abnormal remaining living cells, the living cells may also involve abnormal chromosomal structure, living cells with abnormal number of chromosomes 1 to 22 except for the 5 common abnormal numbers of chromosomes. The cell types are lymphocytes of peripheral blood, umbilical cord blood, and bone marrow.

[0017] 2. After mixing the blood with an equal amount of 1640 culture medium, gradually add it along the tube wall into a centrifuge tube containing 4ml of lymphocyte separation medium (mixed blood: lymphocyte separation medium = 6:4), and centrifuge at 3000 rpm for 10 minutes.

[0018] 3. Absorb the white blood cell layer, transfer it to another centrifuge tube, add 6ml of serum-free 1640 c...

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Abstract

The invention relates to an EBV transfected chimera quality-control cell for antenatal diagnosis and a preparation method thereof, and belongs to medical genetics. The invention is mainly characterized in that the preparation method comprises: taking residual living cells which are used for chromosomal disease diagnosis as needed by clinical diagnosis and treatment and are definitely diagnosed to have common numerical abnormalities of chromosomes, separating lymphocytes, culturing in an RPMI-1640 culture solution containing cyclosporin A and EBV under a condition of 37 DEG C and 5% CO2, allowing B lymphocytes to form immortalized lymphoblast lines with infinite reproduction capability, performing subculture amplification to reach a desired amount, then cryopreserving the cell lines, mixing the cell lines according to a required ratio for quality control when in use so as to prepare each original cell line with only one type of numerical abnormalities of chromosomes into a chimera quality-control cell line which has a specific ratio and multiple combinations of numerical abnormalities of chromosomes, is universal for various testings, and is more difficult for differential diagnosis. Original cells are easily available, and waste excessive cells are prepared artificially into practical and effective quality-control cells which can be amplified as needed.

Description

technical field [0001] The invention relates to an EBV-transfected chimera quality control cell for prenatal diagnosis of birth defects and a preparation method thereof, which is mainly used in cytogenetic laboratories in the medical field for technical assessment, indoor quality control and external quality assessment of abnormal diagnosis of common chromosome numbers . 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 amenorrhea, a...

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Application Information

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IPC IPC(8): C12N5/10C12Q1/70C12Q1/68C12Q1/02
Inventor 翁炳焕
Owner 翁炳焕
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