Therapy and diagnosis of conditions related to telomere length and/or telomerase activity
First Claim
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1. A method for detecting cancer in a human, said method comprising:
- (a) obtaining a cell sample from said individual;
(b) lysing cells in said cell sample to form a cell lysate under conditions such that denaturation of telomerase does not occur;
(c) incubating an aliquot of said cell lysate in a reaction mixture comprising an oligonucleotide primer that can serve as a substrate for telomerase-mediated primer extension, nucleoside triphosphates, and a buffer under conditions such that, if telomerase activity is present, said primer is extended by telomerase-mediated addition of nucleotides derived from said nucleoside triphosphates to said primer;
(d) determining whether said primer has been extended; and
(e) correlating presence of an extended primer with presence of cancer cells in said human and absence of an extended primer with absence of cancer cells in said human.
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Abstract
Method and compositions are provided for the determination of telomere length and telomerase activity, as well as the ability to inhibit telomerase activity in the treatment of proliferative diseases. Particularly, primers are elongated under conditions which minimize interference from other genomic sequences, so as to obtain accurate determinations of telomeric length or telomerase activity. In addition, compositions are provided for intracellular inhibition of telomerase activity.
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Citations
11 Claims
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1. A method for detecting cancer in a human, said method comprising:
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(a) obtaining a cell sample from said individual; (b) lysing cells in said cell sample to form a cell lysate under conditions such that denaturation of telomerase does not occur; (c) incubating an aliquot of said cell lysate in a reaction mixture comprising an oligonucleotide primer that can serve as a substrate for telomerase-mediated primer extension, nucleoside triphosphates, and a buffer under conditions such that, if telomerase activity is present, said primer is extended by telomerase-mediated addition of nucleotides derived from said nucleoside triphosphates to said primer; (d) determining whether said primer has been extended; and (e) correlating presence of an extended primer with presence of cancer cells in said human and absence of an extended primer with absence of cancer cells in said human. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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Specification