Methods of quantifying viral load in an animal with a ribonuclease resistant RNA preparation
AssigneesAmbion, Inc., Cenetron Diagnostics, LLC
InventorsBrittan L. Pasloske, Dwight B. DuBois, David M. Brown, Matthew M. Winkler
Life of this patent
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Abstract
The present invention relates to nuclease resistant nucleic acids in general and ribonuclease resistant RNAs in particular. Methods of making and using such nucleic acids are disclosed.
Claims66
A method of determining the presence or absence of and/or quantifying a viral load of an RNA virus in an animal comprising:
- obtaining a sample from the animal;
- isolating sample RNA from the sample;
- obtaining a ribonuclease resistant standard comprising a recombinant RNA segment standard encapsidated in viral coat protein, wherein the recombinant RNA segment standard comprises a region encoding a sequence that can be a standard for the detection and/or quantification of a virus;
- assaying the RNA isolated from the sample for the presence of a sequence that is indicative of the virus;
- and
- employing the RNA segment standard as a standard to determine the presence or absence of and/or quantify a viral load of the virus in the animal.
A method of determining the presence or absence of and/or quantifying a viral load of HIV in an animal comprising:
- obtaining a sample from the animal;
- obtaining a ribonuclease resistant standard comprising a recombinant RNA segment standard encapsidated in viral coat protein, wherein the recombinant RNA segment standard comprises a region encoding a sequence that can be a standard for the detection and/or quantification of HIV;
- admixing the sample and the standard;
- isolating both sample RNA from the sample and the recombinant RNA segment standard from the viral coat protein;
- assaying the sample RNA for the presence of a sequence that is indicative of HIV;
- and
- employing the RNA segment standard as a standard for the detection and/or quantification of HIV as a standard
- to determine the presence or absence of and/or quantify a viral load of HIV in the animal.
A method of determining the presence or absence of and/or quantifying a viral load of HCV in an animal comprising:
- obtaining a sample from the animal;
- obtaining a ribonuclease resistant standard comprising a recombinant RNA segment standard encapsidated in viral coat protein, wherein the recombinant RNA segment standard comprises a region encoding a sequence that can be a standard for the detection and/or quantification of HCV;
- admixing the sample and the standard;
- isolating both sample RNA from the sample and the recombinant RNA segment standard from the viral coat protein;
- assaying the sample RNA for the presence of a sequence that is indicative of HCV;
- and
- employing the RNA segment standard as a standard for the detection and/or quantification of HCV as a standard
- to determine the presence or absence of and/or quantify a viral load of HCV in the animal.
A method of determining the presence or absence of and/or quantifying a viral load of HTLV in an animal comprising:
- obtaining a sample from the animal;
- obtaining a ribonuclease resistant standard comprising a recombinant RNA segment standard encapsidated in viral coat protein, wherein the recombinant RNA segment standard comprises a region encoding a sequence that can be a standard for the detection and/or quantification of HTLV;
- admixing the sample and the standard;
- isolating both sample RNA from the sample and the recombinant RNA segment standard from the viral coat protein;
- assaying the sample RNA for the presence of a sequence that is indicative of HTLV;
- and
- employing the RNA segment standard as a standard for the detection and/or quantification of HTLV as a standard
- to determine the presence or absence of and/or quantify a viral load of HTLV in the animal.
Dependencies are inferred from the claim text.