Nucleic acid detection in pooled samples
OwnerLance Fors, Bruce P. Neri +3inventors
InventorsLance Fors, Bruce P. Neri, Mary Ann D. Brow, Monika de Arruda Indig, Robert Roeven
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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.
Claims35
A method of performing nucleic acid testing on a pooled sample, comprising:
- a) providing;
- i) a pooled sample, wherein said pooled sample comprises biological material combined from a plurality of individual samples;
- and
- ii) INVADER assay reagents configured to detect the presence or absence of a target nucleic acid sequence;
- and
- b) contacting said pooled sample with said INVADER assay reagents under conditions such that the presence or absence of said target nucleic acid sequence in said pooled sample is determined.
A method of performing nucleic acid testing on a pooled sample, comprising:
- a) providing;
- i) a plurality of individual biological material samples;
- and
- ii) INVADER assay reagents configured to detect the presence or absence of a target nucleic acid sequence;
- and
- b) forming a sub-pool by combining a portion of each of said plurality of individual biological samples, and
- c) contacting said sub-pool with said INVADER assay reagents under conditions such that the presence or absence of said target nucleic acid sequence in said sub-pool is determined.
A method of performing nucleic acid testing on a pooled sample, comprising:
- a) providing;
- i) a pooled sample, wherein said pooled sample comprises biological material combined from a plurality of individual samples;
- and
- ii) INVADER assay reagents configured to detect measure the quantity of a target nucleic acid sequence present in a sample;
- and
- b) contacting said pooled sample with said INVADER assay reagents under conditions such that the quantity of said target nucleic acid sequence present in said pooled sample is determined.
A method for detecting an allele frequency of a polymorphism, comprising:
- a) providing;
- i) a pooled sample, wherein said pooled sample comprises target nucleic acid sequences from a plurality of individuals;
- and
- ii) INVADER assay reagents configured to detect the presence or absence of a polymorphism;
- 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.
A method for detecting an allele frequency of a polymorphism, comprising:
- a) providing;
- i) a pooled sample, wherein said pooled sample comprises target nucleic acid sequences from a plurality of individuals;
- and
- ii) INVADER assay reagents configured to generate distinct signals for each allele of a polymorphic locus in said target nucleic acid sequence;
- b) contacting said pooled sample with said INVADER assay reagents to generate 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.
A method for detecting a rare mutation comprising;
- a) providing;
- i) a sample from a single subject, wherein said sample comprises at least 10,000 target nucleic acid sequences,
- ii) a detection assay capable of detecting a mutation in a population of target nucleic acid sequence that is present at an allele frequency of 1:1000 or less compared to wild type alleles;
- and
- b) assaying said sample with said detection assay under conditions such that the presence or absence of a rare mutation is detected.
A method for detecting a rare mutation comprising;
- a) providing;
- i) a sample from a single subject, wherein said sample comprises at least 10,000 target nucleic acid sequences,
- ii) a detection assay capable of detecting a mutation in a population of target nucleic acid sequence that is present at an allele frequency of 1:1000 or less compared to wild type alleles;
- and
- b) assaying said sample with said detection assay under conditions such that an allele frequency in said sample of a rare mutation is determined.
Dependencies are inferred from the claim text.