Nucleic acid detection in pooled samples
First Claim
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1. A method for detecting an allele frequency of a polymorphism in a target nucleic acid sequence, comprising:
- a) providing;
i) a pooled sample, wherein said pooled sample comprises a plurality of target nucleic acids, each comprising an allele of said target nucleic acid sequence; and
ii) INVADER assay reagents configured to detect the presence or absence of a polymorphism in said target nucleic acid sequence; and
b) contacting said pooled sample with said INVADER assay reagents to generate a detectable signal; and
c) measuring said detectable signal, thereby determining a number of said target nucleic acid sequences that contain said polymorphism, wherein said allele frequency is determined from said number of said target nucleic acid sequences that contain said polymorphism.
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Abstract
The present invention relates to detecting target nucleic acid sequences in pooled samples. In particular, the present invention relates to compositions and methods for detecting the presence or absence of target nucleic acid sequences (e.g. RNA virus sequences) in a pooled sample employing an INVADER detection assay. In certain embodiments, the present invention allows target nucleic acid sequence detection in pooled biological samples (e.g. pooled blood samples) without prior amplification of the target.
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22 Claims
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1. A method for detecting an allele frequency of a polymorphism in a target nucleic acid sequence, comprising:
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a) providing;
i) a pooled sample, wherein said pooled sample comprises a plurality of target nucleic acids, each comprising an allele of said target nucleic acid sequence; and
ii) INVADER assay reagents configured to detect the presence or absence of a polymorphism in said target nucleic acid sequence; and
b) contacting said pooled sample with said INVADER assay reagents to generate a detectable signal; and
c) measuring said detectable signal, thereby determining a number of said target nucleic acid sequences that contain said polymorphism, wherein said allele frequency is determined from said number of said target nucleic acid sequences that contain said polymorphism. - View Dependent Claims (2, 3, 4, 5)
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6. A method for detecting an allele frequency of a polymorphism in a target nucleic acid sequence, comprising:
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a) providing;
i) a pooled sample, wherein said pooled sample comprises a plurality of target nucleic acids, each comprising at least one allele of said target nucleic acid sequence; and
ii) INVADER assay reagents configured to generate distinct signals for each different allele of said target nucleic acids; and
b) contacting said pooled sample with said INVADER assay reagents to generate a at least one distinct signal; and
c) measuring each of said at least one distinct signal, thereby determining a proportion of each allele of said polymorphic locus within said pooled sample, wherein said allele frequency is determined from said proportion. - View Dependent Claims (7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
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Specification