Kit for noninvasively acquiring rare cell of fetus and method of kit

A rare cell and kit technology, applied in the field of biomedicine, can solve the problems of invasive and easy contamination of rare fetal cells

Active Publication Date: 2019-08-13
广州莱德盟生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the shortcomings of the prior art described above, the purpose of the present invention is to provide a kit and method for obtaining fetal r

Method used

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  • Kit for noninvasively acquiring rare cell of fetus and method of kit
  • Kit for noninvasively acquiring rare cell of fetus and method of kit
  • Kit for noninvasively acquiring rare cell of fetus and method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 (verification of the identification efficiency of endoplasmic reticulum probes using cell line models)

[0068] In this example, a trophoblast cell line was used to simulate a clinical sample to observe and verify the identification efficiency of endoplasmic reticulum probes for trophoblast cells from maternal background cells.

[0069] 1. Experimental equipment

[0070] Eppendorf centrifuge, Olympus inverted fluorescence microscope BX51, BD FACSAria flow cytometer.

[0071] 2. Experimental reagents

[0072] HTR8-S / Vneo, JEG-3, and JAR cell lines were purchased from the Cell Bank of the Chinese Academy of Sciences.

[0073] RPMI 1640 Medium (Cat. No.: 11875093), DMEM High Glucose Medium (Cat. No.: 10569044), Fetal Bovine Serum (Cat. No.: 10437028), Trypsin (Cat. No.: 25300054), Phosphate Buffered Saline PBS (Cat. No.: C10010500BT-1) Purchased from Thermo Fisher Scientific Corporation.

[0074] Red blood cell lysate (product number: AR1118) was purchased fr...

Embodiment 2

[0095]Embodiment 2 (acquisition of rare fetal trophoblast cells)

[0096] This example provides a method for rapidly obtaining pure fetal rare cells in a pregnant woman's secretion specimen, including the screening, rapid identification and single cell picking of fetal trophoblast cells. This embodiment utilizes the principle that the trophoblast cells in the birth canal of pregnant women secrete a large amount of proteins such as β-HCG and progesterone, so the endoplasmic reticulum content in the cells is abnormally rich, and can be distinguished from squamous epithelial cells and white blood cells and other maternal cells. By staining samples specifically for the endoplasmic reticulum and scoring accordingly, they can be quickly and easily differentiated from background cells.

[0097] 1. Experimental equipment

[0098] Olympus inverted fluorescence microscope BX51, AVIOS GmbhCellCelector automatic cell separation and screening system, Eppendorf centrifuge.

[0099] 2. Exp...

Embodiment 3

[0119] Example 3 (Whole Genome Amplification and Identification of Rare Fetal Trophoblastic Cells)

[0120] 1. Experimental equipment

[0121] The instruments used include Eppendorf nucleic acid amplification instrument, Nanodrop 2000 nucleic acid detection instrument, Beijing Liuyi electrophoresis instrument, and BioRad gel exposure instrument. The capillary electrophoresis detection used by STR was completed by Shanghai Yihe Applied Biotechnology Co., Ltd.; Sanger sequencing was completed by Sangon Bioengineering (Shanghai) Co., Ltd.

[0122] 2. Experimental reagents

[0123] Single-cell whole genome amplification kit (article number: KT110700150) was purchased from Jiangsu Yikang Gene Technology Co., Ltd.; 21 STR Detection Kit (Cat. No.: DC8902) was purchased from Promega Biotechnology Co., Ltd.; Hot Start Ex Taq PCR Kit (Cat. No.: RR006Q), 100bp DNA Maker (Cat. No.: 3422A), 6×loading Buffer (Product No.: 9156) was purchased from Takara Co., Ltd.; primers were synthes...

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Abstract

The invention provides a kit for noninvasively acquiring a rare cell of a fetus and a method of the kit. The method comprises the following steps that 1) cells are provided; 2) an endoplasmic reticulum fluorescence probe and different colors of cell fluorochromes of the endoplasmic reticulum fluorescence probe are adopted for dyeing the collected cells; 3) according to the fluorescence intensity difference between a target cell and background cells, trophoblast cells of the fetus are obtained through quick screening; 4) the target cell is obtained through picking of a cell sorting platform; 5)the obtained target cell is used for antenatal diagnosis of single-gene diseases. The new method for acquiring the fetal cell from secreta of a pregnant woman is created for the first time. The operation steps are simple, the consumed time is short, the number of the acquired cells is stable, the purity of the cells is high, and the acquired trophoblast cells of the fetus have integral fetal genomes; on the basis of low cost, quite pure fetal genomes are acquired, the complete noninvasive purpose is achieved, and the method is suitable for antenatal diagnosis of alpha-thalassemia, beta-thalassemia and other single-gene diseases.

Description

technical field [0001] The invention relates to the field of biomedical technology, in particular to a kit for non-invasively obtaining fetal rare cells and a method thereof. Background technique [0002] Thalassemia, thalassemia, thalassemia, also known as thalassemia, is a group of genetic hemolytic diseases caused by a defect in the globin gene, resulting in reduced or absent globin chain synthesis. A group of disabling and fatal genetic blood diseases that seriously threaten health. Thalassemia is the most widely distributed and accumulated autosomal recessive genetic disease in the world. According to the type of inhibited globin peptide chain synthesis, it can be divided into α-thalassemia, β-thalassemia, γ-thalassemia, δ-thalassemia, δβ-thalassemia, etc., among which α-thalassemia and β-thalassemia Most common. In southern China, the incidence of thalassemia is very high. The incidence of α-thalassemia can reach 3.18-9.46% and 9.47-14.13% in Guangdong and Guangxi r...

Claims

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Application Information

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IPC IPC(8): C12N5/073C12Q1/02
CPCC12N5/0603G01N33/5005C12N2509/00
Inventor 郑磊黄一芳司徒博叶昕怡江秀娟
Owner 广州莱德盟生物科技有限公司
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