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.
Claims25
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 oligonucleotide primers, a first primer which comprises 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:6 and SEQ ID NO:7, and a second primer which comprises at least 10 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:8 and SEQ ID NO:9;
- and
- c) detecting the amplification product using a labeled probe, 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 which comprises at least 10 consecutive nucleotides of a sequence according to SEQ ID NO:11, and a second primer which comprises at least 10 consecutive nucleotides of a sequence according to SEQ ID NO:12;
- and
- c) detecting or genotyping the amplification product using one or more probes.
A kit for the detection or genotyping of dengue virus RNA in a sample, comprising two oligonucleotide primers, a first primer being about 15-26 nucleotides in length and comprising at least 10 consecutive nucleotides of a sequence according to SEQ ID NO:11, and a second primer being about 15-26 nucleotides in length and comprising at least 10 consectutive nucleotides of a sequence according to SEQ ID NO:12, and at least one probe.
A kit for the detection of dengue virus type 2 RNA in a sample, said kit comprising two oligonucleotide primers, a first primer being about 15-26 nucleotides in length and 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:6 and SEQ ID NO:7, and a second primer being about 15-26 nucleotides in length and comprising at least 10 consecutive nucleotides of a sequence 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.
An oligonucleotide of about 15-26 nucleotides in length comprising at least 10 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:23.
An oligonucleotide of about 15-26 nucleotides in length, comprising at least 10 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NO:11, SEQ ID NO:12, and SEQ ID NO:14.
An oligonucleotide of 15-26 nucleotides in length which is a subsequence of Dengue virus and which comprises at least 15 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:7, and SEQ ID NO:8, wherein said consecutive nucleotides are identical throughout their length to a Dengue virus genome.
An oligonucleotide of 15-26 nucleotides in length which is a subsequence of Dengue virus and which comprises at least 15 consecutive nucleotides of a sequence selected from the group consisting of SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:16, and SEQ ID NO:17, wherein said consecutive nucleotides are identical throughout their length to a Dengue virus genome.
Dependencies are inferred from the claim text.