Claims
- 1. A method of detecting the presence or absence of a fetal DNA sequence of interest in fetal DNA derived from a sample of peripheral blood obtained from a pregnant woman, comprising:
obtaining a sample of peripheral blood from a pregnant woman; treating the sample of peripheral blood such that fetal DNA present in fetal nucleated cells present in the sample is made available for detection resulting in available fetal DNA; and detecting the presence or absence of a fetal DNA sequence of interest in the available fetal DNA.
- 2. The method as claimed in claim 1 wherein the proportion of fetal nucleated cells present in the sample of peripheral blood obtained from the pregnant woman is increased forming a sample enriched in fetal nucleated cells prior to the detection step.
- 3. The method of claim 1 wherein the sample enriched in fetal nucleated cells is formed by separating the non-nucleated cells from the nucleated in the peripheral blood sample forming a nucleated cell enriched sample and by treating the nucleated cell enriched sample such that the proportion of fetal cells in the sample is increased forming a fetal nucleated cell enriched sample.
- 4. The method of claim 1 wherein the fetal DNA is amplified prior to the detection step resulting in amplified fetal DNA.
- 5. A method of detecting the presence or absence of fetal DNA of interest in fetal DNA derived from a sample of peripheral blood obtained from a pregnant woman, comprising:
obtaining a sample of peripheral blood from the woman; treating the sample of peripheral blood such that fetal DNA present in fetal nucleated cells present in the sample is made available for hybridization with a DNA probe resulting in available fetal DNA; contacting the available fetal DNA with a DNA probe hybridizable to fetal DNA of interest under hybridization conditions; and detecting the presence or absence of hybridization between the DNA probe and the fetal DNA of interest as an indication of the presence or absence of the fetal DNA of interest.
- 6. The method of claim 5 wherein the fetal nucleated cells in the treating step are undifferentiated hematopoietic cells.
- 7. The method of claim 5 wherein the fetal nucleated cells are selected from the group consisting of erythroblasts, lymphoblasts, and myeloblasts.
- 8. The method of claim 6 wherein the undifferentiated hematopoietic cells are erythroid cells.
- 9. The method of claim 8 wherein the undifferentiated erythroid cells are fetal nucleated erythrocytes.
- 10. The method of claim 5 wherein the proportion of fetal nucleated cells present in the sample of peripheral blood obtained from the pregnant woman is increased forming a sample enriched in fetal nucleated cells prior to the detection step.
- 11. The method of claim 10 wherein the sample enriched in fetal nucleated cells is formed by separating non-nucleated cells from nucleated cells in the peripheral blood sample forming a nucleated cell enriched sample and by treating the nucleated cell enriched sample such that the proportion of fetal cells in the sample is increased forming a fetal nucleated cell enriched sample.
- 12. The method of claim 5 wherein the fetal DNA of interest in the contacting and detecting steps is Y chromosomal DNA.
- 13. The method of claim 5 wherein the hybridization in the detecting step is between the DNA probe and a disease causing mutation.
- 14. The method of claim 13 wherein the disease causing mutation is a cystic fibrosis-causing mutation.
- 15. The method of claim 13 wherein the disease causing mutation is a Duchenne muscular dystrophy-causing mutation.
- 16. The method of claim 13 wherein the disease causing mutation is a hemophilia A-causing mutation.
- 17. The method of claim 13 wherein the disease causing mutation is a Gaucher disease-causing mutation.
- 18. The method of claim 13 wherein the disease causing mutation is a sickle cell anemia-causing mutation.
- 19. The method of claim 13 wherein the hybridization in the detecting step is between the DNA probe and a fetal DNA of interest selected for assessing a chromosomal abnormality.
- 20. The method of claim 5 wherein the hybridization in the detecting step is in situ hybridization.
- 21. The method of claim 6 wherein the hybridization in the detecting step is in situ hybridization.
- 22. The method of claim 9 wherein the hybridization in the detecting step is in situ hybridization.
- 23. The method of claim 12 wherein the hybridization in the contacting step is in situ hybridization.
- 24. The method of claim 5 wherein the fetal DNA is amplified prior to the detection step resulting in amplified fetal DNA.
- 25. The method of claim 6 wherein the fetal DNA is amplified prior to the detection step resulting in amplified fetal DNA.
- 26. The method of claim 9 wherein the fetal DNA is amplified prior to the detection step resulting in amplified fetal DNA.
- 27. The method of claim 3 further comprising determining the gestational age at the time the sample of peripheral blood is obtained and treating the sample such that fetal DNA present in a selected type of fetal nucleated cells is made available for hybridization, said type of fetal nucleated cell being selected based upon the known presence of such fetal nucleated cells in the peripheral blood of a pregnant woman at the determined gestational age.
- 28. The method of claim 5 further comprising the step of treating the peripheral blood sample to remove residual fetal nucleated cells from a previous pregnancy prior to the detection step.
- 29. The method of claim 5 wherein the peripheral blood sample in the obtaining step is obtained between about ten and about nineteen weeks gestation.
- 30. A method of detecting a fetal DNA sequence of interest in fetal DNA derived from a sample of peripheral blood obtained from a pregnant woman, comprising:
obtaining a sample of peripheral blood from a pregnant woman; separating fetal nucleated cells from the peripheral blood onto a solid support forming immobilized fetal nucleated material; contacting the immobilized fetal nucleated material with a DNA probe hybridizable to fetal DNA of interest under hybridization conditions; and detecting the presence of hybridization between the DNA probe and the fetal DNA of interest as an indication of the presence or absence of the fetal DNA of interest.
- 31. The method of claim 30 wherein the hybridization in the detecting step is in situ hybridization.
- 32. The method of claim 31 wherein the hybridization of high copy number repeat sequences present in the fetal DNA is suppressed in the contacting step.
- 33. A method for determining the sex of a fetus, comprising:
obtaining a sample of peripheral blood from a woman pregnant with a fetus; treating the sample of peripheral blood such that fetal DNA present in fetal nucleated cells present in the sample is made available for hybridization resulting in available fetal DNA; contacting the available fetal DNA with a DNA probe hybridizable to fetal Y chromosomal DNA under hybridization conditions; and detecting the presence of hybridization between the DNA probe and the fetal Y chromosomal DNA as an indication of a male fetua or the absence of hybridization as an indication of a female fetus.
- 34. A method for diagnosing a disease in a fetus, comprising:
obtaining a sample of peripheral blood from a woman pregnant with a fetus; treating the sample of peripheral blood such that fetal DNA present in fetal nucleated cells present in the peripheral sample is made available for hybridization resulting in available fetal DNA; contacting the available DNA with a DNA probe hybridizable to fetal DNA of interest associated with a disease under hybridization conditions; and detecting the presence or absence of hybridization between the DNA probe and the fetal DNA of interest associated with the disease as an indication that the fetus has the disease.
- 35. A method for detecting a chromosomal abnormality in a fetus, comprising:
obtaining a sample of peripheral blood from a woman pregnant with a fetus; separating fetal nucleated cells from the peripheral blood sample onto a solid support forming immobilized fetal nucleated material; contacting the immobilized fetal nucleated materail with a DNA probe hybridizable to chromosomal fetal DNA of interest under hybridization conditions; and detecting the presence or absence of hybridization between the DNA probe and the chromosomal fetal DNA of interest as an indication of the presence or absence of a chromosomal abnormality.
- 36. The method of claim 35 wherein the hybridization in the detecting step is in situ hybridization.
- 37. The method of claim 36 wherein the chromosomal abnormality is a chromosomal aneuploidy.
- 38. The method of claim 37 wherein the chromosomal aneuploidy is trisomy 21.
- 39. The method of claim 37 wherein the chromosomal aneuploidy is trisomy 18.
- 40. The method of claim 37 wherein the chromosomal aneuploidy is trisomy 13.
- 41. A method for determining whether a pregnancy is at risk, comprising:
obtaining a peripheral blood sample from a pregnant woman at a selected gestational age; detecting the number of fetal cells present in the peripheral blood sample; comparing the number of fetal cells detected to a standard to determine whether the number of fetal cells is indicative of a pregnancy at risk, said standard being selected based on the number of fetal cells present in a peripheral blood sample obtained from a woman having a normal pregnancy at the selected gestational age.
- 42. A method of increasing the proportion of fetal nucleated cells present in a peripheral blood sample containing nucleated and non-nucleated cells, comprising:
separating non-nucleated cells from nucleated cells present in a peripheral blood sample forming a nucleated cell enriched sample; and treating the nucleated cell enriched sample such that the proportion of fetal cells in the sample is increased forming a fetal nucleated cell enriched sample.
- 43. The method of claim 42 wherein the separating step is a density gradient centrifugation.
- 44. The method of claim 42 wherein the nucleated enriched sample is treated by contacting the sample with (i) a first monoclonal antibody which recognizes fetal nucleated cells but not maternal cells and/or (ii) a second monoclonal antibody which recognizes maternal cells but not fetal nucleated cells, under conditions appropriate for antibody binding thereby producing (i) fetal nucleated cell-first monoclonal antibody complexes and/or (ii) maternal cell-second monoclonal antibody complexes, respectively, and
(i) separating fetal nucleated cell-first monoclonal antibody complexes from maternal cells and/or (ii) separating maternal cell-second monoclonal antibody complexes from fetal nucleated cells thereby separating fetal nucleated cells from maternal cells.
- 45. The method of claim 43 wherein the nucleated enriched sample is treated by contacting the sample with (i) a first monoclonal antibody which recognizes fetal nucleated cells but not maternal cells and/or (ii) a second monoclonal antibody which recognizes maternal cells but not fetal nucleated cells, under conditions appropriate for antibody binding producing (i) fetal nulceated cell-first monoclonal antibody complexes and/or (ii) maternal cell-second monoclonal antibody complexes, respectively and,
(i) separating fetal nucleated cell-first monoclonal antibody complexes from maternal cells and/or (ii) separating maternal cell-second monoclonal antibody complexes from fetal nucleated cells thereby separating fetal nucleated cells from maternal cells.
RELATED APPLICATIONS
[0001] This application is a continuation-in-part application of U.S. Ser. No. 07/772,689, filed on Oct. 7, 1991, which is a continuation-in-part application of U.S. Ser. No. 07/706,393, filed on May 28, 1991, now abandoned, which is a continuation-in-part of U.S. Ser. No. 07/436,057, filed on Nov. 13, 1989, now abandoned, all being entitled “Non-Invasive Method for Isolation and Detection of Fetal DNA” by Diana W. Bianchi. The contents of all of the forementioned applications are hereby expressly incorporated by reference.
Continuations (5)
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08470633 |
Jun 1995 |
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09827853 |
Apr 2001 |
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08338279 |
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08470633 |
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07957736 |
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Continuation in Parts (3)
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07772689 |
Oct 1991 |
US |
Child |
07957736 |
Oct 1992 |
US |
Parent |
07706393 |
May 1991 |
US |
Child |
07772689 |
Oct 1991 |
US |
Parent |
07436057 |
Nov 1989 |
US |
Child |
07706393 |
May 1991 |
US |