P
USH1825HExpiredUtilityPatentIndex 57

Isothermal transcription based assay for the detection of HTLV I and HTLV II RNA

Assignee: AKZO NOBEL NVPriority: Jan 30, 1998Filed: Jan 30, 1998Granted: Dec 7, 1999
Est. expiryJan 30, 2018(expired)· nominal 20-yr term from priority
Inventors:ROMANO JOSEPH WSHURTLIFF ROXANNEWILLIAMS KIMBERLY G
C12Q 1/702C12Q 2531/143
57
PatentIndex Score
4
Cited by
35
References
21
Claims

Abstract

An isothermal transcription based amplification assay for the detection of HTLV I RNA uses primers and probes for sequences within the tax gene, the pol gene, or the gag gene. An isothermal transcription based amplification assay for the detection of HTLV II uses primers and probes for sequences within the tax gene. Target specific primers and probes are also disclosed.

Claims

exact text as granted — not AI-modified
We claim: 
     
       1. An oligonucleotide of 10-50 nucleotides, comprising at least 10 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NO:1 , SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:27, and SEQ ID NO:28, 
     
     
       2. An oligonucleotide comprising the oligonucleotide of claim 1 further comprising the sequence of SEQ ID NO:29 at the 5' end thereof. 
     
     
       3. A method for the detection of HTLV I RNA in a sample, comprising: a) obtaining a sample which may contain HTLV I RNA;   b) performing isothermal transcription based amplification on the sample with 2 oligonucleotide primers, one of which comprises at least 10 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:15, and SEQ ID NO:16, and the other of which comprises at least 10 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:6, SEQ ID NO:10, and SEQ ID NO:14; and   c) detecting the resulting product of step b), whereby detection of the amplification product indicates the presence of HTLV I RNA in the sample.   
     
     
       4. The method of claim 3, wherein the sample comprises cells and RNA is extracted from the cells in the sample prior to step b). 
     
     
       5. The method of claim 3, wherein the detection step uses a labeled probe, wherein the probe comprises a sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13 and SEQ ID NO:17, whereby hybridization of the probe to the amplification product indicates the presence of HTLV I RNA in the sample. 
     
     
       6. A kit for the detection or quantitation of HTLV I RNA in a sample, comprising two primers selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:1, SEQ ID NO:6, SEQ ID NO:10, and SEQ ID NO:14. 
     
     
       7. The kit of claim 6 wherein the primers are SEQ ID NO:1 and SEQ ID NO:3. 
     
     
       8. The kit of claim 6 wherein the primers are SEQ ID NO:6 and SEQ ID NO:8. 
     
     
       9. The kit of claim 6 wherein the primers are SEQ ID NO:10 and SEQ ID NO:12. 
     
     
       10. The kit of claim 6 wherein the primers are SEQ ID NO:14 and SEQ ID NO:16. 
     
     
       11. The kit of claim 6 further comprising, disposed in a separate container, a probe comprising a sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13 and SEQ ID NO:17. 
     
     
       12. The kit of claim 6 wherein the primers are SEQ ID NO:18 and SEQ ID NO:23. 
     
     
       13. The kit of claim 6 wherein the primers are SEQ ID NO:19 and SEQ ID NO:23. 
     
     
       14. The kit of claim 6 wherein the primers are SEQ ID NO:26 and SEQ ID NO:27. 
     
     
       15. A method for the detection of HTLV II RNA in a sample, comprising: a) obtaining a sample which may contain HTLV II RNA;   b) performing isothermal transcription based amplification on the sample with 2 oligonucleotide primers, one of which comprises at least 10 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:25, and SEQ ID NO:26 and the other of which comprises at least 10 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NO:18, SEQ ID NO:19, and SEQ ID NO:27; and   c) detecting the resulting product of step b), whereby detection of the amplification product indicates the presence of HTLV II RNA in the sample.   
     
     
       16. The method of claim 15, wherein the sample comprises cells and RNA is extracted from the cells in the sample prior to step b). 
     
     
       17. The method of claim 15, wherein the detection step uses a labeled probe, wherein the probe comprises a sequence selected from the group consisting of SEQ ID NO:24 and SEQ ID NO:28, whereby hybridization of the probe to the amplification product indicates the presence of HTLV II RNA in the sample. 
     
     
       18. A kit for the detection or quantitation of HTLV II RNA in a sample, comprising two primers selected from the group consisting of SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:26 SEQ ID NO:18, SEQ ID NO:19, and SEQ ID NO:27. 
     
     
       19. The kit of claim 18 wherein the primers are SEQ ID NO:18 and SEQ ID NO:21. 
     
     
       20. The kit of claim 18 wherein the primers are SEQ ID NO:19 and SEQ ID NO:21. 
     
     
       21. The kit of claim 18 further comprising, disposed in a separate container, a probe comprising a sequence selected from the group consisting of SEQ ID NO:24 and SEQ ID NO:28.

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