Detection of microsatellite instability and its use in diagnosis of tumors
View Patent ↗Methods and kits are disclosed for use in the analysis of microsatellite instability in genomic DNA. Methods and kits are also disclosed which can be used to detect microsatellite instability DNA present in biological materials, such as tumors. The methods and kits of the present invention can be used to detect or diagnose diseases associated with microsatellite instability, such as certain types of cancer.
1. A method of analyzing microsatellite loci, comprising:
a) co-amplifying at least three quasi-monomorphic mononucleotide repeat loci of human genomic DNA from at least one sample of genomic DNA in a multiplex amplification reaction comprising one or more primers, thereby producing amplified DNA fragments; and
b) determining the size of the amplified DNA fragments to detect the presence or absence of microsatellite instability.
2. The method of claim 1 , wherein at least one quasi-monomorphic repeat loci is selected from the group consisting of MONO-11, MONO-15, BAT-25, and BAT-26.
3. The method of claim 1 , wherein at least one quasi-monomorphic repeat loci comprises a perfect mononucleotide repeat comprising greater than 25 adenine residues.
4. The method of claim 1 , wherein at least one quasi-monomorphic mononucleotide repeat loci comprises from 25 to 35 nucleotides.
5. The method of claim 1 , wherein the at least one sample of genomic DNA comprises a first sample of genomic DNA originating from normal non-cancerous biological material from an individual and a second sample of genomic DNA originating from a tumor of the individual the method further comprising: detecting microsatellite instability by comparing the size of the amplified DNA fragments produced from co-amplifying the first sample of genomic DNA to the size of the amplified DNA fragments produced from co-amplifying the second sample of genomic DNA.
6. The method of claim 1 , wherein the at least one sample of genomic DNA is from a tumor, further comprising evaluating microsatellite instability by comparing the fragment sizes of step (b) to the most commonly observed allele sizes for the at least three quasi-monomorphic mononucleotide repeat loci in a population.