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US 11,884,969 B2Granted
Isothermal amplification components and processes
AssigneeNat Diagnostics, Inc.
InventorsAndrew P. Miller, Honghua Zhang
Life of this patent
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Abstract
The technology relates in part to methods and compositions for isothermal amplification of nucleic acids.
Claims30
A method for detecting a target nucleic acid sequence in a sample, the method comprising:
- (a) amplifying a target nucleic acid sequence in a sample under an isothermal amplification condition, wherein the target nucleic acid sequence comprises a first strand and a second strand complementary to each other, and wherein the amplifying comprises contacting a double-stranded nucleic acid comprising the target nucleic acid sequence with:
- (i) a first primer and a second primer, wherein the first primer is capable of hybridizing to a sequence of the first strand of the target nucleic acid sequence, and the second primer is capable to hybridizing to a sequence of the second strand of the target nucleic acid sequence;
- and
- (ii) an enzyme having a hyperthermophile polymerase activity, thereby generating a nucleic acid amplification product, wherein the nucleic acid amplification product comprises:
- (1) the sequence of the first primer or a portion thereof, or the reverse complement thereof,
- (2) the sequence of the second primer or a portion thereof, or the reverse complement thereof, and
- (3) a spacer sequence flanked by (1) and (2), wherein the spacer sequence is 1 to 10 bases long;
- and
- (b) detecting the nucleic acid amplification product, wherein the detecting is performed in 20 minutes or less from the time the double-stranded nucleic acid is contacted with (a)(i) the first and second primers and (a)(ii) the enzyme having a hyperthermophile polymerase activity,
- wherein the method does not comprise using any enzymes that are not a polymerase.
7 more elements
Dependencies are inferred from the claim text.