Replication Stable and RNase Resistant Chimeras of Pestivirus with Insertion in 3' Nontranslated Region (3'NTR)
First Claim
1. A chimeric viral RNA sequence comprising:
- first virus RNA sequence from Pestivirus, and second RNA sequence inserted within a 3′
NTR of the first Pestivirus RNA sequence, wherein the chimeric viral RNA sequence is stable in RNA replication and forms a ribonuclease resistant viral particle.
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Abstract
The invention relates to the field of nucleic acid amplification, particularly to quality control materials for use in viral RNA assays. It specifically relates to the construction of a recombinant Pestivirus by the identification of a region in the 3′NTR of the viral RNA genome where additional sequence elements can be stably inserted. Chimeric Pestivirus with sequence insertions in the 3′ nontranslated region (3′NTR) of the viral RNA genome were stable in replication and capable of forming infectious, RNase resistant virus particles. This chimeric Pestivirus with a 3′NTR insertion can be utilized as a quality control material in analytical assays for RNA targets, including external, internal controls, quantitative standards in PCR and NAT nucleic acid assays.
25 Citations
25 Claims
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1. A chimeric viral RNA sequence comprising:
- first virus RNA sequence from Pestivirus, and second RNA sequence inserted within a 3′
NTR of the first Pestivirus RNA sequence, wherein the chimeric viral RNA sequence is stable in RNA replication and forms a ribonuclease resistant viral particle. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
- first virus RNA sequence from Pestivirus, and second RNA sequence inserted within a 3′
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12. A vector comprising:
- DNA sequence of a chimeric viral RNA sequence comprising;
vector DNA sequence, first DNA sequence derived from a first Pestivirus RNA sequence, and second RNA sequence inserted within a 3′
NTR of the first Pestivirus RNA sequence, wherein the sequence of the chimeric RNA virus derived from the DNA vector is stable in replication, and forms a RNase resistant viral particle. - View Dependent Claims (13, 14, 15, 16, 17)
- DNA sequence of a chimeric viral RNA sequence comprising;
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18. A culture of cells containing RNA virus comprising:
- host cell harboring chimeric first and second RNA virus sequences, wherein the first RNA virus is Pestivirus and a second RNA sequence is inserted in a 3′
NTR of the first Pestivirus virus RNA sequence, wherein the chimeric RNA viral sequence is stable in RNA replication, and ribonuclease resistant viral particles comprising the first and second RNA virus sequences when cultured. - View Dependent Claims (19, 20, 21, 22, 23, 24, 25)
- host cell harboring chimeric first and second RNA virus sequences, wherein the first RNA virus is Pestivirus and a second RNA sequence is inserted in a 3′
Specification