Disease detection by digital protein truncation assays
AssigneeThe Johns Hopkins University
InventorsC. Giovanni Traverso, Kenneth W. Kinzler, Bert Vogelstein
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Abstract
Genetic diseases can be diagnosed by detection of mutations in causative genes. Protein truncation assays can be used to detect gene products of truncation-type mutations. However, the sensitivity of the assays is often insufficient to detect mutations present in a sample of DNA at a low frequency. Sensitivity can be increased by dividing samples so that the signal generated by a mutant allele comprises a larger fraction of the total alleles than prior to dividing. Thus a previously undetectable signal generated by the mutant allele can become detectable in the assay. Such increased sensitivity permits detection at early stages and in samples having high levels of other alleles.
Claims61
A method of detecting tumors, comprising:
- dividing a test sample of APC alleles isolated from a patient, to form a plurality of aliquots of APC alleles;
- amplifying said APC alleles in said plurality of aliquots to form amplified APC alleles;
- transcribing and translating proteins in vitro using said amplified APC alleles as transcription templates;
- determining size or composition of said proteins, wherein proteins which differ in size or composition from the protein produced by a wild-type APC allele indicate a mutation in an amplified APC allele which indicates a tumor in the patient.
A method of detecting a disease associated with a mutation in a gene, comprising:
- dividing a test sample of alleles of the gene isolated from a patient, to form a plurality of aliquots of alleles of the gene;
- amplifying said alleles in said plurality of aliquots to form amplified alleles;
- transcribing and translating proteins in vitro using said amplified alleles as transcription templates;
- determining size or composition of said proteins, wherein proteins which differ in size or composition from the protein produced by a wild-type allele of the gene indicate a mutation in an amplified allele of the gene which indicates the disease in the patient.
Dependencies are inferred from the claim text.