Nucleic acid-based tests for RHD typing, gender determination and nucleic acid quantification

• US 8,173,370 B2
• Filed: 02/07/2008
• Issued: 05/08/2012
• Est. Priority Date: 02/08/2007
• Status: Expired due to Fees

• Current Assignee: Sequenom Incorporated (Laboratory Corporation Of America Holdings)
• Original Assignee: Sequenom Incorporated (Laboratory Corporation Of America Holdings)
• Inventors: Oeth, Paul A., Ehrich, Mathias, Lee, Min S.
• Primary Examiner(s): Kim, Young J
• Application Number:
  US12/027,954
• Publication Number:
  US 20080299562A1
• Time in Patent Office: 1,552 Days
• Field of Search: None
• US Class Current: 435/6.12
• CPC Class Codes: C12Q 1/6881 for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays

Abstract

The invention in part provides nucleic acid-based assays, which are particularly useful for non-invasive prenatal testing. The invention in part provides compositions and methods for RhD typing, detecting the presence of fetal nucleic in a sample, determining the relative amount of fetal nucleic acid in a sample and determining the sex of a fetus, wherein each of the assays may be performed alone or in combination.

Claims

1. A multiplex method for determining the presence or absence of one or more RhD exons in a fetal nucleic acid, comprising:

simultaneously amplifying fetal nucleic acid from a pregnant female with two or more amplification primers yielding amplification products,

contacting the amplification products with extension primers under extension conditions, thereby generating extension products, and

determining the presence or absence of an RhD exon in the fetal nucleic acid based on the extension products;

wherein;

the extension primers independently comprise a nucleotide sequence of;

2. The method of claim 1, wherein the fetal nucleic acid is from the blood of the pregnant female.

3. The method of claim 1, wherein the fetal nucleic acid is from serum from the pregnant female.

4. The method of claim 1, wherein the fetal nucleic acid is from plasma from the pregnant female.

5. The method of claim 1, wherein the presence or absence of a 37 base pair insertion in exon 4 of RhD is determined.

6. The method of claim 5, wherein the zygosity of a 37 base pair insertion in exon 4 of RhD is determined.

7. The method of claim 1, wherein the presence or absence of exon 4 of RhD is determined.

8. The method of claim 1, wherein the presence or absence of exon 5 of RhD is determined.

9. The method of claim 1, wherein the presence or absence of exon 7 of RhD is determined.