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) amplifying one or more quasi-monomorphic mononucleotide repeat loci of human genomic DNA from at least one sample of human genomic DNA in an amplification reaction comprising one or more primers, thereby producing at least one amplified DNA fragment, wherein at least one mononucleotide repeat locus comprises MONO-11; and
b) determining the size of the amplified DNA fragments.
2. The method of claim 1 , wherein the one or more quasi-monomorphic mononucleotide repeat loci further comprises MONO-15.
3. A method of analyzing microsatellite loci, comprising:
a) co-amplifying at least two mononucleotide repeat loci of human genomic DNA from at least one sample of human genomic DNA in a multiplex amplification reaction comprising primers, thereby producing amplified DNA fragments, wherein at least one mononucleotide repeat locus is selected from the group consisting of MONO-11 and MONO-15; and
b) determining the size of the amplified DNA fragments.
4. The method of claim 3 , wherein the mononucleotide repeat loci comprise MONO-11 and MONO-15.
5. The method of claim 3 , wherein the mononucleotide repeat loci comprise at least one mononucleotide repeat locus in addition to MONO-11 or MONO-15.
6. The method of claim 5 , wherein the additional mononucleotide repeat locus comprises at least one mononucleotide repeat locus selected from the group consisting of BAT-25, BAT-26, BAT-40, and TGFBR-II.
7. The method of claim 1 , wherein at least three mononucleotide repeat loci are co-amplified.
8. The method of claim 7 , wherein the at least three mononucleotide repeat loci include a set of mononucleotide repeat loci selected from the group consisting of:
MONO-11 and BAT-25; MONO-11 and BAT-26; MONO-11 and BAT-40; MONO-11 and TGFBR-II; MONO-11, BAT-25, and BAT-26; MONO-11, BAT-25, and BAT-40;
MONO-11, BAT-25, and TGFBR-II; MONO-11, BAT-26, and BAT-40; MONO-11, BAT-26, and TGFBR-II; MONO-11, BAT-40, and TGFBR-II; MONO-15 and BAT-25;
MONO-15 and BAT-26; MONO-15 and BAT-40; MONO-15 and TGFBR-II; MONO-15, BAT-25, and BAT-26; MONO-15, BAT-25, and BAT-40; MONO-15, BAT-25, and
TGFBR-II; MONO-15, BAT-26, and BAT-40; MONO-15, BAT-26, and TGFBR-II; and MONO-15, BAT-40, and TGFBR-II.
9. A method of analyzing microsatellite loci, comprising steps of:
a) providing primers for co-amplifying a set of at least three microsatellite loci of human genomic DNA, comprising at least one mononucleotide repeat locus and at least two tetranucleotide repeat loci, wherein the at least one mononucleotide repeat locus comprises MONO-11;
b) co-amplifying the set of at least three micro satellite loci from at least one sample of genomic DNA in a multiplex amplification reaction, using the primers, thereby producing amplified DNA fragments; and
c) determining the size of the amplified DNA fragments.