Isothermal transcription based assay for the detection and genotyping of dengue virus
AssigneeAkzo Nobel N.V.
InventorsJoseph W. Romano, Eun Mi Lee, Gregory J. Hurteau
Life of this patent
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Abstract
An isothermal transcription based amplification assay for dengue virus RNA uses primer combinations for sequences within the envelope gene or the 3' non-coding region of the virus and a probe. Probes may be specific for a serotype of dengue virus.
Claims8
A method for the detection of dengue virus RNA, comprising:
- a) obtaining a sample which may contain dengue virus RNA;
- b) performing an isothermal transcription based amplification on the sample with two primers, a first primer selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, and SEQ ID NO:21, and a second primer selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:8and SEQ ID NO:9;
- and
- c) detecting the amplification product using a labeled probe, wherein the probe is selected from the group consisiting of SEQ ID NO:5 and SEQ ID NO:10, whereby hybridization of the probe to the amplification product indicates the presence of dengue virus RNA in the sample.
A method for the detection or genotyping of dengue virus RNA in a sample comprising:
- a) obtaining a sample which may contain dengue virus RNA;
- b) performing an isothermal transcription based amplification on the sample with two primers, a first primer selected from the group consisting of SEQ ID NO:11 and SEQ ID NO:22, and a second primer containing SEQ ID NO:12;
- and
- c) detecting the amplification product using a probe, wherein the probe is selected from the group consisting of SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16 and SEQ ID NO:17, whereby hybridization of the amplification product with SEQ ID NO:13 indicates the virus RNA is type 1, hybridization of the amplification product with SEQ ID NO:14 indicates the virus RNA is type 2, hybridization of the amplification product with SEQ ID NO:15 indicates the virus RNA is type 3, hybridization of the amplification product with SEQ ID NO:16 indicates the virus RNA is type 4, and wherein hybridization of the amplification product with SEQ ID NO:17 indicates the presence of dengue virus RNA.
A kit for the detection or genotyping of dengue virus RNA in a sample, comprising two primers, a first primer containing an oligonucleotide selected from the group consisting of SEQ ID NO:11 and SEQ ID NO:22, and a second primer containing SEQ ID NO:12, and at least one probe, wherein the at least one probe is selected from the group consisting of SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16 and SEQ ID NO:17.
A kit for the detection of dengue virus type 2 RNA in a sample, said kit comprising two primers, a first primer containing an oligonucleptide selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:20, and SEQ ID NO:21, and a second primer containing an oligonucleotide selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:8, and SEQ ID NO:9;
- and a probe, wherein the probe is selected from the group consisting of SEQ ID NO:5, SEQ ID NO:10, and SEQ ID NO:23.
An oligonucleotide selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:20, SEQ ID NO:
- 21, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:23.
An oligonucleotide selected from the group consisting of SEQ ID NO:
- 11, SEQ ID NO:20, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, and SEQ ID NO:17.
Dependencies are inferred from the claim text.