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hTERT and SV40 transduced quality-control cell for molecular genetics diagnosis and preparation method thereof

A genetic and cellular technology, applied in the field of molecular genetic diagnosis of hTERT and SV40 transduction quality control cells and their preparation, which can solve problems such as lack of quality control substances

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

[0010] In order to solve the problem that there is no quality control substance in the diagnosis of chromosomal diseases in the medical molecular genetics diagnosis laboratory, the inventor proposes the present invention

Method used

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  • hTERT and SV40 transduced quality-control cell for molecular genetics diagnosis and preparation method thereof
  • hTERT and SV40 transduced quality-control cell for molecular genetics diagnosis and preparation method thereof

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Embodiment Construction

[0018] 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 that grow adherently, and 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.

[0019] 2. Extraction of SV40LT and hTERT: (1) Enzyme digestion of SV40LT and hTERT: ① Enzyme digestion of SV40LT: 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, 18uL H 2 O and restriction endonuclease BamH I (1-5...

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Abstract

The invention relates to an hTERT and SV40 transduced quality-control cell for molecular genetics diagnosis and a preparation method thereof. The invention is mainly characterized in that the preparation method comprises: connecting recons of SV40LTag-pcDNA3.1(-) and pLXSNneo-hTERT by T4DNA, introducing tissue cells with common numerical abnormalities of chromosomes by liposome, screening cells integrated with the recons by G418, performing subculture amplification and cryopreservation, mixing the cells according to a required ratio for quality control when in use so as to convert each original cell with only one type of numerical abnormalities of chromosomes into a chimera quality-control cell with a certain ratio of 2-5 types of numerical abnormalities of chromosomes. Therefore, the difficulty for differential diagnosis and chimera diagnosis is increased; the original cells are easily available; waste residual cells are converted into effective immortalized quality-control cells; difficult quality-control cells are used for external quality assessment and internal quality control; and the quality-control cell of the invention has important significance on in-time discovery and solution of quality problems, and diagnostic level improvement.

Description

technical field [0001] The invention relates to a molecular genetic diagnosis hTERT and SV40 transduction quality control cell and a preparation method thereof, which are mainly used in medical molecular genetic diagnosis laboratories for technical assessment of chromosomal disease diagnosis, indoor quality control and external quality evaluation. 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, and hermaphroditism. Chro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10
Inventor 翁炳焕
Owner 翁炳焕
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