Method for evaluating microsatellite instability in a tumor sample
OwnerBharati Bapat, Melanie Anne Roseinventors
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Abstract
A detection method for high throughput screening for tumor microsatellite instability. The method employs a panel of microsatellite loci and it is based on a fluorescent multiplex PCR system. The method provides a fast, sensitive, and cost-effective high throughput screening method of MSI detection. The method allows many samples to be processed in one day on a single polyacrylamide gel, and it utilizes much less nucleic acid sample than conventional methods.
Claims20
A method for evaluating microsatellite instability in a tumor sample by detecting microsatellite loci in the sample comprising:
- (a) forming a polymerase chain reaction mixture comprising the tumor sample, a polymerase and primer sets for at least two selected microsatellite loci associated with cancer, each primer set characterized by (a) a forward primer containing a sequence complimentary to a 5′ upstream primer-specific portion of a selected microsatellite loci;
- and (b) a reverse primer complementary to a 3′ downstream primer-specific portion of the same microsatelite loci, wherein one of the primers has a detectable reporter label;
- (b) subjecting the polymerase chain reaction mixture to polymerase chain reaction cycles to form amplified products complementary to microsatellite loci sequences in the tumor sample;
- (c) detecting the reporter labels and distinguishing the amplified products to indicate the presence of one or more of the microsatellite loci in the sample;
- and
- (d) repeating steps (a) to (c) with primer sets for at least two different selected microsatellite loci.
A method for diagnosing colorectal cancer or hereditary non-polyposis colorectal cancer syndrome in a human individual comprising the steps of (a) isolating DNA from the human individual;
- (b) assaying the DNA using multiplex polymerase chain reaction for microsatellite loci associated with colorectal cancer or hereditary non-polyposis colorectal cancer syndrome relative to a normal human individual (c) diagnosing colorectal cancer or hereditary non-polyposis colorectal cancer syndrome in the human individual based on the frequency of microsatellite loci.
Dependencies are inferred from the claim text.