COMPOSITIONS FOR USE IN IDENTIFICATION OF HERPESVIRUSES
First Claim
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1. A composition, comprising at least two oligonucleotide primer pairs that are selected from the group consisting of:
- at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A20G23C24T22] base composition from a Herpes simplex 1 virus nucleic acid, a [A19G23C16T21] base composition from a Human herpesvirus 2 nucleic acid, and a [A18G12C25T24] base composition from a Bovine herpesvirus 2 nucleic acid;
at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A16G19C17T21] base composition from a Herpes simplex 1 virus nucleic acid, a [A16G18C19T20] base composition from a Human herpesvirus 2 nucleic acid, and a [A15G17C18T23] base composition from a Bovine herpesvirus 2 nucleic acid;
at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A24G43C36T24] base composition from a Herpes simplex 1 virus nucleic acid, a [A26G34C41T26] base composition from a Human herpesvirus 2 nucleic acid, and a [A26G34C41T26] base composition from a Bovine herpesvirus 2 nucleic acid;
at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A19G31C25T17] base composition from a Herpesvirus aotus nucleic acid and a [A16G9C25T21] base composition from a Human herpesvirus 5 nucleic acid;
at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A16G23C14T22] base composition from a Herpesvirus aotus nucleic acid, a Human herpesvirus 1 nucleic acid, a Human herpesvirus 4 nucleic acid, and a Human herpesvirus 6 nucleic acid;
at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A15G34C19T33] base composition from a Human herpesvirus 1 nucleic acid and a Human herpesvirus 6 nucleic acid;
at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A12G22C14T13] base composition from a Bovine herpesvirus 2 nucleic acid and a Human herpesvirus 1 nucleic acid, a [A12G23C14T12] base composition from a Human herpesvirus 2 nucleic acid, and a [A13G20C18T10] base composition from a Human herpesvirus 4 nucleic acid; and
at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A17G17C24T13] base composition from a Herpes simplex 1 virus nucleic acid, a Human herpesvirus 4 nucleic acid, and a Human herpesvirus 5 nucleic acid.
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Abstract
The present invention relates generally to identification of herpesviruses and provides methods, compositions and kits useful for this purpose when combined, for example, with molecular mass or base composition analysis.
99 Citations
38 Claims
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1. A composition, comprising at least two oligonucleotide primer pairs that are selected from the group consisting of:
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at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A20G23C24T22] base composition from a Herpes simplex 1 virus nucleic acid, a [A19G23C16T21] base composition from a Human herpesvirus 2 nucleic acid, and a [A18G12C25T24] base composition from a Bovine herpesvirus 2 nucleic acid; at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A16G19C17T21] base composition from a Herpes simplex 1 virus nucleic acid, a [A16G18C19T20] base composition from a Human herpesvirus 2 nucleic acid, and a [A15G17C18T23] base composition from a Bovine herpesvirus 2 nucleic acid; at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A24G43C36T24] base composition from a Herpes simplex 1 virus nucleic acid, a [A26G34C41T26] base composition from a Human herpesvirus 2 nucleic acid, and a [A26G34C41T26] base composition from a Bovine herpesvirus 2 nucleic acid; at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A19G31C25T17] base composition from a Herpesvirus aotus nucleic acid and a [A16G9C25T21] base composition from a Human herpesvirus 5 nucleic acid; at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A16G23C14T22] base composition from a Herpesvirus aotus nucleic acid, a Human herpesvirus 1 nucleic acid, a Human herpesvirus 4 nucleic acid, and a Human herpesvirus 6 nucleic acid; at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A15G34C19T33] base composition from a Human herpesvirus 1 nucleic acid and a Human herpesvirus 6 nucleic acid; at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A12G22C14T13] base composition from a Bovine herpesvirus 2 nucleic acid and a Human herpesvirus 1 nucleic acid, a [A12G23C14T12] base composition from a Human herpesvirus 2 nucleic acid, and a [A13G20C18T10] base composition from a Human herpesvirus 4 nucleic acid; and at least one primer pair comprising sequences that are configured to generate amplicons comprising a [A17G17C24T13] base composition from a Herpes simplex 1 virus nucleic acid, a Human herpesvirus 4 nucleic acid, and a Human herpesvirus 5 nucleic acid. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33)
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- 2. A purified oligonucleotide primer pair for identifying a herpesvirus in a sample, said primer pair comprising a forward primer and a reverse primer, each configured to hybridize to nucleic acid of two or more different herpesvirus species or strains in a nucleic acid amplification reaction which produces an amplification product between about 29 to about 200 nucleobases in length, said amplification product comprising portions corresponding to a forward primer hybridization region, a reverse primer hybridization region and an intervening region having a base composition which varies among amplification products produced from nucleic acid of said two or more different herpesvirus species or strains, said base composition of said intervening region providing a means for identifying said herpesvirus.
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14. An isolated amplification product for identification of a herpesvirus, said amplification product produced by a process comprising:
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a) amplifying nucleic acid of a herpesvirus in a reaction mixture comprising a primer pair, said primer pair comprising a forward primer and a reverse primer, each configured to hybridize to nucleic acid of two or more different herpesviruses in a nucleic acid amplification reaction, said amplification product having a length of about 29 to about 200 nucleobases and comprising portions corresponding to a forward primer hybridization region, a reverse primer hybridization region and an intervening region having a base composition which varies among amplification products produced from nucleic acid of said two or more different herpesviruses, said base composition of said intervening region providing a means for identifying said herpesvirus; and b) isolating said amplification product from said reaction mixture. - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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34. A kit comprising one or more purified primer pairs for identifying a herpesvirus in a sample, each member of said one or more primer pairs having at least 70% sequence identity with a corresponding member of one or more primer pairs selected from the group consisting of:
- SEQ ID NOs;
36;
11, 59;
60, 32;
57, 33;
1, 10;
23, 58;
4, 2;
44 and 29;
9. - View Dependent Claims (35, 36)
- SEQ ID NOs;
Specification