Kit for detection of telomerase reverse transcriptase nucleic acids
View Patent ↗The invention provides compositions and methods related to human telomerase reverse transcriptase (hTRT), the catalytic protein subunit of human telomerase. The polynucleotides and polypeptides of the invention are useful for diagnosis, prognosis and treatment of human diseases, for changing the proliferative capacity of cells and organisms, and for identification and screening of compounds and treatments useful for treatment of diseases such as cancers.
1. A kit for detecting a nucleic acid that encodes human telomerase reverse transcriptase (hTRT), or fragment thereof, in a sample, the kit comprising a pair of amplification primers, said pair comprising a first amplification primer at least 10 nucleotides in length that hybridizes specifically to a DNA having the sequence of SEQ. ID NO: 1 or the complement thereof at 5° C. to 25° C. below T m in aqueous solution at 1 M NaCl but under the same reaction conditions does not hybridize to a DNA having the sequence of SEQ. ID NO:8 or the complement thereof; and a second amplification primer at least 10 nucleotides in length that hybridizes specifically to a DNA having the sequence of SEQ. ID NO: 1 or the complement thereof at 5° C. to 25° C. below T m in aqueous solution at 1 M NaCl.
2. The kit of claim 1 wherein the both the first and the second amplification primers hybridize specifically to a DNA having the sequence of SEQ. ID NO: 1 or the complement thereof but do not hybridize to a DNA having the sequence of SEQ. ID NO:8 or the complement thereof.
3. A kit for detecting a nucleic acid that encodes human telomerase reverse transcriptase (hTRT), or fragment thereof, in a sample, the kit comprising a pair of oligonucleotide amplification primers, said pair comprising a first amplification primer that comprises at least 10 consecutive nucleotides of SEQ. ID NO:1 or the complement thereof that are not contained in SEQ. ID NO:8 or its complement; and a second amplification primer that comprises at least 10 consecutive nucleotides of SEQ. ID NO:1 or the complement thereof.
4. The kit of claim 3 , wherein at least one of said primers comprises at least 20 consecutive nucleotides of SEQ. ID NO: 1 or the complement thereof.
5. The kit of claim 3 , wherein both of said primers comprise at least 20 consecutive nucleotides of SEQ. ID NO: 1 or the complement thereof.
6. The kit of claim 3 , wherein each of said primers comprise 15 to 30 consecutive nucleotides of SEQ. ID NO: 1 or the complement thereof.
7. The kit of claim 3 , comprising a plurality of nested sets of primers.
8. The kit of claim 3 , further comprising one or more labeled probes.
9. The kit of claim 3 , further containing reagents for isolating mRNA.
10. The kit of claim 3 , further comprising a DNA polymerase and a reagent comprising deoxyribonucleoside triphosphates (dNTPs).
11. The kit of claim 3 , further comprising instructional materials for conducting an assay to detect a nucleic acid that encodes human telomerase reverse transcriptase (hTRT), or fragment thereof.
12. The kit of claim 3 , which is a kit for diagnosis or prognosis of cancer.
13. The kit of claim 3 wherein the both the first and the second amplification primers comprise at least 10 consecutive nucleotides of SEQ. ID NO:1 or the complement thereof that are not contained in SEQ. ID NO:8 or its complement.
14. A pair of oligonucleotide primers for amplifying a nucleic acid that encodes human telomerase reverse transcriptase (hTRT) or fragment thereof, wherein each primer comprises at least 10 consecutive nucleotides of SEQ. ID NO: 1 or the complement thereof, and at least one primer comprises at least 10 consecutive nucleotides of SEQ ID NO:1 or the complement thereof that are not contained in SEQ. ID NO:8 or the complement thereof.
15. The primers of claim 14 , each of which comprises at least 20 consecutive nucleotides of SEQ. ID NO: 1 or the complement thereof.
16. The primers of claim 14 , each of which comprises 15 to 30 consecutive nucleotides of SEQ. ID NO: 1 or the complement thereof.