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Method of identifying foetal erythroblast

An erythroblastic, fetal technique applied in the field of identifying and/or isolating at least one fetal nucleated primitive red blood cell, identifying at least one fetal nucleated primitive red blood cell in a sample

Inactive Publication Date: 2015-05-06
NAT UNIV OF SINGAPORE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thus, it is not inconceivable that a very small number of fetal cells (~20 cells) enriched from blood from a mother undergoing a euploid pregnancy might actually be sufficient for non-invasive prenatal diagnosis

Method used

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  • Method of identifying foetal erythroblast
  • Method of identifying foetal erythroblast
  • Method of identifying foetal erythroblast

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Materials and methods

[0084] Intensive literature studies have been conducted on the presence and functional role of plasma membrane proteins characteristic of FPNRBCs in various human tissues and cells, including the fetus (trophoblast / placenta). A brief description of the location of these proteins, their physiological roles (including those associated with human fetal development), and diseases associated with their mutations has been provided above, along with available data on similar functions of AARBCs.

[0085] FPNRBCs can be isolated from WBCs in maternal blood by negative depletion of CD45-positive cells, and these ideal cells for non-invasive prenatal diagnosis can be enriched by AARBCs if known suitable surface antigens for FPNRBCs are available. Membrane proteins of FPNRBCs were graphically represented by mass spectrometry data, and the data graphs were compared with data graphs of AARBCs membrane proteomes known in the art to identify specific surface me...

Embodiment 2

[0211] Enrichment of FPNRBCs from post-termination of pregnancy (TOP) maternal blood using anti-ASCT2 antibody following a three-step enrichment protocol

[0212] Maternal blood after 10 mls TOP was collected from two patients. Blood samples were processed using our laboratory's three-step enrichment protocol. Briefly, diluted blood samples were layered on Percoll 1118 density medium and centrifuged. The interface was collected and all leukocytes were depleted by magnetic active cell sorting (MACS) using anti-CD45 magnetic beads. Cells from the negative fraction were incubated with anti-ASCT2 antibody for 30 minutes, washed, and then incubated with anti-rabbit IgG-magnetic beads for indirect MACS (positive) selection of FPNRBCs. 20 FPNRBCs were able to be recovered from each sample (Table 11).

[0213] Table 11 - Enrichment of FPNRBCs from maternal blood after TOP using anti-ASCT2 antibody

[0214] Maternal blood after TOP

gestational age of the fetus

Volu...

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Abstract

There is provided a method for identifying at least one foetal erythroblast the method comprising: (a) detecting the expression of at least one foetal erythroblast specific marker selected from the group consisting of neutral amino acid transporter B (SLC1A5), solute carrier family 3 (activators of dibasic and neutral amino acid transport) member 2 isoform A (SLC3A2), Splice Isoform A of Chloride channel protein 6, Transferrin receptor protein 1, Splice Isoform 3 of Protein GPR107 precursor, Olfactory receptor 11H4, Splice Isoform 1 of Protein C9orf5, Cleft lip and palate transmembrane protein 1, BCG induced integral membrane protein BIGM103, Antibacterial protein FALL-39 precursor, CAAX prenyl protease 1 homolog, Splice Isoform 2 of Synaptophysin-like protein, Vitamin K epoxide reductase complex subunit 1-like protein 1, Splice Isoform 1 of Protein C20orf22 (ABHD12), Hypothetical protein DKFZp564K247 (Hypoxia induced gene 1 protein) (IPI Accession No. IPI00295621), Hypothetical protein DK-FZp586C1924 (IPI Accession No. IPI00031064), ALEX3 protein variant, Hypothetical protein MGC14288 (IPI Accession No. IPI00176708), protein with IPI Accession No. IPI00639803 and protein with IPI Accession No. IPI00646289, wherein detection of the marker indicates the presence of the foetal erythroblast.

Description

technical field [0001] The present invention generally relates to methods for identifying and / or isolating at least one fetal nucleated erythrocyte primitive. In particular, the present invention relates to a method for identifying at least one fetal nucleated red blood cell primitive in a sample by detecting at least one membrane protein specific for the fetal nucleated red blood cell primitive. Background technique [0002] Currently, prenatal diagnosis of chromosomal and monogenic disorders relies on invasive procedures for cytogenetic and / or molecular analysis (eg, amniocentesis, chorionic villus sampling (CVS), or fetal blood sampling (FBS)) to obtain fetal cells. These invasive tests carry a small but significant risk of aborting the fetus. On the one hand it limits the understanding of diagnostic tests due to the fear of losing the fetus, while on the other hand it leads to the death of an otherwise healthy fetus. [0003] Non-invasive methods of diagnosing the gen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53C07K16/28C40B30/04C12Q1/68
CPCC12N5/0603C12N5/0641G01N33/56966G01N2800/385G01N2800/387C12Q1/6883C12Q1/6888C12Q2600/158C07K16/28C07K14/705C12Q2600/156
Inventor 马海施·楚兰尼速酷马·颇努萨米张火明林青松安妮沙·布蒂里·马幼淋普里亚·卡当派
Owner NAT UNIV OF SINGAPORE
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