Detection of synthetic oligonucleotides extracted from body fluids or tissues
First Claim
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1. A method of detecting synthetic oligonucleotides that are present in body fluids or tissues, said oligonucleotides being extractable with isobutanol and ethyl ether, the method comprising the steps of:
- (a) proteolytically digesting a body fluid or tissue sample;
(b) extracting the proteolytically digested sample with a partitioning agent;
(c) extracting the aqueous phase in the extracted sample with isobutanol and ethyl ether,(d) drying the remaining solution containing nucleic acids to a pellet and resuspending the nucleic acids into a solution;
(e) binding the nucleic acids to a hybridization membrane;
(f) treating the membrane having bound nucleic acids with a labeled oligonucleotide probe that is complementary to and specifically binds to the synthetic oligonucleotide to be detected;
(g) washing the membrane to remove any labeled oligonucleotide that is not specifically hybridized to nucleic acids bound to the membrane; and
(h) detecting the presence of the probe on the membrane.
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Abstract
The invention provides a method for detecting specific synthetic oligonucleotides that are present in body fluid or tissue samples taken from an animal or human patient to whom oligonucleotides have been administered.
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Citations
12 Claims
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1. A method of detecting synthetic oligonucleotides that are present in body fluids or tissues, said oligonucleotides being extractable with isobutanol and ethyl ether, the method comprising the steps of:
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(a) proteolytically digesting a body fluid or tissue sample; (b) extracting the proteolytically digested sample with a partitioning agent; (c) extracting the aqueous phase in the extracted sample with isobutanol and ethyl ether, (d) drying the remaining solution containing nucleic acids to a pellet and resuspending the nucleic acids into a solution; (e) binding the nucleic acids to a hybridization membrane; (f) treating the membrane having bound nucleic acids with a labeled oligonucleotide probe that is complementary to and specifically binds to the synthetic oligonucleotide to be detected; (g) washing the membrane to remove any labeled oligonucleotide that is not specifically hybridized to nucleic acids bound to the membrane; and (h) detecting the presence of the probe on the membrane. - View Dependent Claims (2, 3, 4, 5, 6)
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7. A method of detecting the presence in body fluids or tissues of a synthetic oligonucleotide, the method comprising detecting the presence on a membrane of an oligonucleotide duplex formed by a labeled oligonucleotide probe hybridized to the synthetic oligonucleotide to be detected, said synthetic oligonucleotide capable of being extracted by isobutanol and ethyl ether, the oligonucleotide duplex on the membrane having been prepared by:
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(a) proteolytically digesting a body fluid or tissue sample; (b) extracting the proteolytically digested sample with a partitioning agent; (c) extracting the aqueous phase in the extracted sample with isobutanol and ethyl ether; (d) drying the remaining solution containing nucleic acids to a pellet and resuspending the nucleic acids into a solution; (e) binding the nucleic acids to a hybridization membrane; (f) treating the membrane having bound nucleic acids with a labeled oligonucleotide probe that is complementary to and specifically binds to the synthetic oligonucleotide to be detected; (g) washing the membrane to remove any labeled oligonucleotide that is not specifically hybridized to nucleic acids bound to the membrane. - View Dependent Claims (8, 9, 10, 11, 12)
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Specification