Non-invasive prenatal diagnosis

• US 6,258,540 B1
• Filed: 11/29/1999
• Issued: 07/10/2001
• Est. Priority Date: 03/04/1997
• Status: Expired due to Term

• Current Assignee: Oxford University Innovation Limited (University of Oxford)
• Original Assignee: Isis Innovation Limited
• Inventors: Lo, Yuk-Ming Dennis, Wainscoat, James Stephen
• Primary Examiner(s): Arthur, Lisa B.
• Assistant Examiner(s): GOLDBERG, JEANINE ANNE
• Application Number:
  US09/380,696
• Time in Patent Office: 589 Days
• Field of Search: 435/6, 435/91.2, 435/440, 435/91.5
• US Class Current: 435/6.12
• CPC Class Codes: C12Q 1/6879 for sex determination
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers

Abstract

The invention relates to a detection method performed on a maternal serum or plasma sample from a pregnant female, which method comprises detecting the presence of a nucleic acid of foetal origin in the sample. The invention enables non-invasive prenatal diagnosis including for example sex determination, blood typing and other genotyping, and detection of pre-eclampsia in the mother.

Claims

1. A method for detecting a paternally inherited nucleic acid of fetal origin performed on a maternal serum or plasma sample from a pregnant female, which method comprises

amplifying a paternally inherited nucleic acid from the serum or plasma sample and

detecting the presence of a paternally inherited nucleic acid of fetal origin in the sample.

2. The method according to claim 1, wherein the foetal nucleic acid is amplified by the polymerase chain reaction.

3. The method according to claim 2, wherein at least one foetal sequence specific oligonucleotide primer is used in the amplification.

4. The method according to claim 1, wherein the foetal nucleic acid is detected by means of a sequence specific probe.

5. The method according to claim 1, wherein the presence of a foetal nucleic acid sequence from the Y chromosome is detected.

6. The method according to claim 5, wherein the Y chromosome sequence is from the DYS14 locus.

7. The method according to claim 5, wherein the Y chromosome sequence is from the SRY gene.

8. The method according to claim 1, wherein the presence of a foetal nucleic acid from a paternally-inherited non-Y chromosome is detected.

9. The method according to claim 8, wherein the non-Y sequence is a blood group antigen gene.

10. The method according to claim 8, wherein the non-Y sequence is a gene which confers a disease phenotype in the foetus.

11. The method according to claim 8, for Rhesus D genotyping a foetus in a Rhesus D negative mother.

12. The method according to claim 5, for determining the sex of the foetus.

13. The method according to claim 5, which comprises determining the concentration of the foetal nucleic acid sequence in the maternal serum or plasma.

14. The method according to claim 13, wherein the determination of the concentration of foetal nucleic acid sequence in the maternal serum or plasma is by quantitative PCR.

15. The method according to claim 13, for the detection of a maternal or foetal condition in which the level of foetal DNA in the maternal serum or plasma is higher or lower than normal.

16. The method according to claim 13, wherein the pattern of variation of foetal DNA concentration in the maternal serum or plasma at particular stages of gestation is different from normal.

17. The method according to claim 13, for detection of pre-eclampsia.

18. The method according to claim 13, for detection of a foetal chromosomal aneuploidy.

19. The method according to claim 1, wherein the sample contains foetal DNA at a fractional concentration of total DNA of at least about 0.14%, without subjecting it to a foetal DNA enrichment step.

20. The method according to claim 19, wherein the fractional concentration of foetal DNA is at least about 0.39%.

21. A method of performing a prenatal diagnosis, which method comprises the steps of:

(i) providing a maternal blood sample;

(ii) separating the sample into a cellular and a non-cellular fraction;

(iii) detecting the presence of a nucleic acid of foetal origin in the non-cellular fraction according to the method of claim 1;

(iv) providing a diagnosis based on the presence and/or quantity and/or sequence of the foetal nucleic acid.

22. The method according to claim 21, wherein the non-cellular fraction as used in step (iii) is a plasma fraction.

23. The method according to claim 21, including performing the further step of allowing clotting in the maternal sample and using the resulting serum in step (iii).

26. The method according to claim 9, wherein the blood group antigen gene is the rhesus D gene.

27. The method according to claim 10, wherein the gene is the rhesus D gene.

24. A method for detecting a paternally inherited nucleic acid on a maternal blood sample, which method comprises:

removing all or substantially all nucleated and anucleated cell populations from the blood sample,

amplifying a paternally inherited nucleic acid from the remaining fluid and subjecting the amplified nucleic acid to a test for the Paternally inherited fetal nucleic acid.

25. A method for performing a prenatal diagnosis on a maternal blood sample, which method comprises

obtaining a non-cellular fraction of the blood sample

amplifying a paternally inherited nucleic acid from the non-cellular fraction

and performing nucleic acid analysis on the amplified nucleic acid to detect paternally inherited fetal nucleic acid.