Enrichment and identification of fetal cells in maternal blood and ligands for such use

A technology of fetal cells and maternal blood, applied in biochemical equipment and methods, microbial determination/examination, instruments, etc., can solve the problem of not identifying fetal nucleated red blood cells

Active Publication Date: 2013-08-21
アルセディバイオテックエーピーエス
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is suggested that this antibody can be used to recognize maternal white blood cells, but not fetal nucleated red blood cells

Method used

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  • Enrichment and identification of fetal cells in maternal blood and ligands for such use
  • Enrichment and identification of fetal cells in maternal blood and ligands for such use
  • Enrichment and identification of fetal cells in maternal blood and ligands for such use

Examples

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example 1

[0324] Blood sample preparation

[0325] 24 mL peripheral blood samples were obtained from pregnant women at 11 to 14 weeks' gestation. The blood samples were drawn before a non-invasive procedure and after consent was obtained. All blood samples were collected in heparinized tubes and processed immediately after their collection.

[0326] In addition to the heparinized blood, 5 ml of blood was drawn into EDTA tubes. This blood is used for fetal sex analysis. Fetal sex was determined by real-time PCR of cell-free fetal DNA using Y-chromosome-specific genes. Only blood samples from male pregnancy were further processed.

[0327] fixed

[0328] Use pre-coated pipettes (pre-coating buffer is 2% BSA in PBS, w / o Ca 2+ and Mg 2+ ), aliquot 3 mL of whole blood per sample into pre-coated 50 mL centrifuge tubes (8 tubes per sample). Add two mL of 10% formaldehyde in PBS to each tube using a pre-coated pipette. After careful mixing, the blood was fixed at room temperature for 1...

example 2

[0349] Whole blood selection and in-column staining

[0350] blood collection

[0351] Peripheral blood samples of 30 ml were obtained from pregnant women at 11 to 14 weeks of gestation. The blood samples were drawn before a non-invasive procedure and after consent was obtained. All blood samples were collected in heparinized or EDTA tubes and processed within 4 hours of their collection.

[0352] In addition to the heparinized blood, draw 5 mL of blood into EDTA tubes. This blood is used for fetal sex analysis. Fetal sex was determined by real-time PCR of cell-free fetal DNA using Y-chromosome-specific genes.

[0353] Preparation of blood samples - CD105 selection

[0354] 20 to 50 µl of CD105 microbeads (Miltenyi) were added per ml of blood, and after mixing the samples, they were incubated at room temperature for 30 minutes. After incubation, blood samples were aliquoted into six precoated 50 mL tubes (precoating buffer was 2% BSA in PBS w / o Ca 2+ and Mg 2+ ), and 2...

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Abstract

The present invention relates to enrichment and / or identification of fetal cells of a maternal blood sample using fetal cell specific ligands and / or fetal cell specific hybridization probes wherein the ligand or probes are directed to an endothelial / mesenchymal marker, e.g. CD105, CD146 or CD141, in a first round of enrichment and the ligand or probes, in a second round of enrichment, are directed to an epithelial marker, e.g. a cytokeratin, such as CK7, CK8, CK18 or CK19. Enriched or identified fetal cells may be subjected to steps of detection or diagnosis, wherefore the present invention enables non- invasive 5 prenatal diagnostics.

Description

Background technique [0001] Many pregnant women have fetal cell testing for early detection of fetal diseases and genetic abnormalities, especially when the mother is of advanced age (35 years or older) or there is a known genetic disorder in the family. Fetal cells can be obtained by amniocentesis, which removes amniotic fluid from the amniotic cavity in the amniotic sac, or by chorionic villus biopsy, which takes a biopsy from the placenta, so-called invasive sampling. [0002] Prenatal aneuploidy screening uses traditional chromosomal analysis or chromosome-specific DNA probes to illustrate the most frequently abnormal chromosomes in the fetus, specifically the first Aberrations in the number of chromosomes 13, 18, 21, X and Y. [0003] Due to the invasiveness and risk of miscarriage of the aforementioned sampling methods, it would be advantageous to perform fetal diagnosis by a non-invasive procedure, such as the use of maternal blood samples. [0004] During pregnancy, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/569G01N33/68
CPCC12Q1/6806C12Q1/6811C12Q1/6881G01N33/56966G01N2333/4742G01N2333/70596C12Q2600/156G01N21/6486G01N33/6893
Inventor 安德烈亚斯·埃克尔特布里塔·克里斯滕森斯蒂恩·克拉韦奥玛丽·布林克里皮达曼·辛格洛特·哈特
Owner アルセディバイオテックエーピーエス
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