Methods and compositions for identifying a fetal cell
First Claim
1. A method for identifying a fnRBC comprising detecting transcript or protein expression of a HBE, AFP, AHSG, or J42-4-d gene.
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Abstract
The present invention provides methods and compositions for specifically identifying a fetal cell. An initial screening of approximately 400 candidate genes by digital PCR in different fetal and adult tissues identified a subset of 24 gene markers specific for fetal nucleated RBC and trophoblasts. The specific expression of those genes was further evaluated and verified in more defined tissues and isolated cells through quantitative RT-PCR using custom Taqman probes specific for each gene. A subset of fetal cell specific markers (FCM) was tested and validated by RNA fluorescent in situ hybridization (FISH) in blood samples from non-pregnant women, and pre-termination and post-termination pregnant women. Applications of these gene markers include, but are not limited to, distinguishing a fetal cell from a maternal cell for fetal cell identification and genetic diagnosis, identifying circulating fetal cell types in maternal blood, purifying or enriching one or more fetal cells, and enumerating one or more fetal cells during fetal cell enrichment.
230 Citations
31 Claims
- 1. A method for identifying a fnRBC comprising detecting transcript or protein expression of a HBE, AFP, AHSG, or J42-4-d gene.
- 3. A method for identifying a trophoblast comprising detecting transcript or protein expression of a KISS1, LOC90625, AFP, hPL, beta-hCG, or FN1 gene.
- 5. A method for identifying a fetal cell in a maternal sample comprising detecting transcript or protein expression by a cell of one or more of the KISS1, LOC90625, FN1, or AHSG genes to distinguish said fetal cell from a maternal cell.
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6. A method for identifying a fetal cell in a maternal sample comprising detecting transcript or protein expression by a cell of three or more of the hPL, KISS1, LOC90625, FN1, PSG9, HBE, AFP, beta-hCG, AHSG or J42-4-d genes to distinguish said fetal cell from a maternal cell.
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16. A method for identifying a fetal cell in a maternal sample, comprising:
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a. enriching a fetal cell, and b. detecting protein or transcript expression of one or more genes by said fetal cell, wherein said expression of said one or more genes distinguishes said fetal cell from a maternal cell, wherein said one or more genes is hPL, CHS2, KISS1, GDF15, CRH, TFP12, CGB, LOC90625, FN1, COL1A2, PSG9, PSG1, AFP, APOC3, SERPINC1, AMBP, CPB2, ITIH1, APOH, HPX, beta-hCG, AHSG, APOB, or J42-4-d. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28)
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29. A method for identifying a fetal cell specific transcript comprising
a. isolating a transcript from a sample containing a fetal cell and a transcript from a sample lacking fetal cells; -
b. producing cDNAs of said transcripts; c. performing quantitative PCR on said cDNAs; and d. comparing results of said quantitative PCR between samples to identify a marker transcript with higher expression in a fetal cell relative to a non-fetal cell. - View Dependent Claims (30, 31)
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Specification