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US 10,240,214 B2Granted
Methods for discriminating between HIV-1 and lentiviral vectors
AssigneesCalimmune, Inc., Calimmune Australia Pty Ltd, St. Vincent's Hospital Sydney Limited, NewSouth Innovations Pty Limited
InventorsAnthony Dominic Kelleher, Kazuo Suzuki, Geoffrey Phillip Symonds
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Abstract
The present disclosure provides compositions (i.e., amplification primers and probes), methods, and kits that are particularly useful for detecting and/or quantifying nucleic acids present in a sample, such as those derived from HIV or a lentiviral vector.
Claims10
A method of detecting an amount of a lentiviral nucleic acid and an amount of an HIV nucleic acid in a sample comprising:
- (a) contacting the sample with a first forward primer and a first reverse primer;
- (b) contacting the sample with a junction probe specific for a junction site within the 3′ LTR of the lentiviral nucleic acid, wherein the junction probe comprises a first portion which is capable of hybridizing to at least a portion of a sequence within the U3 region of the lentiviral nucleic acid 3′ LTR and a second portion which is capable of hybridizing to at least a portion of a sequence within the R region of the lentiviral nucleic acid 3′ LTR, and wherein the junction probe comprises a first detectable moiety;
- (c) detecting signals from the first detectable moiety;
- (d) contacting the sample with a second probe specific for a TATA-box sequence within a 3′ LTR of an HIV nucleic acid sequence, the second probe conjugated to a second detectable moiety;
- and (e) detecting signals from the second detectable moiety, wherein the detection of the amount of the lentiviral nucleic acid and the amount of the HIV nucleic acid in the sample takes place in the same reaction tube.
2 more elements
A kit comprising:
- (a) a junction probe comprising
- (i) a first region which hybridizes to a sequence within the U3 region of lentiviral nucleic acid 3′ LTR;
- and
- (ii) a second region which hybridizes to a sequence within the R region of lentiviral nucleic acid 3′ LTR;
- and
- (b) a second probe which hybridizes to a TATA-box sequence within 3′ LTR of an HIV nucleic acid;
- wherein the first probe and/or the second probe comprises a detectable label.
4 more elements
Dependencies are inferred from the claim text.