Detection of microsatellite instability and its use in diagnosis of tumors

Abstract

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.

We claim:  
     
         1 . A method of analyzing micro-satellite loci, comprising steps of: 
 a) providing primers for co-amplifying a set of at least three microsatellite loci of genomic DNA, comprising at least one mononucleotide repeat locus selected from the group consisting of BAT-25, BAT-26, BAT-40, MONO-11, and MONO-15, and at least two tetranucleotide repeat loci selected from the group consisting of FGA, D1S518, D1S547, D1S1677, D2S1790, D3S2432, D5S818, D5S2849, D6S1053, D7S3046, D7S1808, D7S3070, D8S1179, D9S2169, D10S1426, D10S2470, D12S391, D17S1294, D17S1299, and D18S51;    b) co-amplifying the set of at least three microsatellite 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.    
     
     
         2 . The method of  claim 1 , wherein the genomic DNA is human genomic DNA.  
     
     
         3 . The method of  claim 1 , wherein the set of at least three microsatellite loci is a set of at least five microsatellite loci, comprising two mononucleotide repeat loci selected from the group consisting of BAT-25, BAT-26, BAT-40, MONO-11, and MONO-15, and three tetranucleotide repeat loci selected from the group consisting of FGA, D1S518, D1S547, D1S1677, D2S1790, D3S2432, D5S818, D5S2849, D6S1053, D7S3046, D7S1808, D7S3070, D8S1179, D9S2169, D10S1426, D10S2470, D12S391, D17S1294, D17S1299, and D18S51.  
     
     
         4 . The method of  claim 1 , wherein at least one of the primers provided in step (a) has a nucleic acid sequence selected from the group of primer sequences identified by: SEQ ID NO: 1-68.  
     
     
         5 . The method of  claim 1 , wherein the set of at least three microsatellite loci is a set of at least nine microsatellite loci, comprising: BAT-25, BAT-26, MONO-15, BAT-40, D3S2432, D7S3046, D7S3070, D7S1808, and D10S1426.  
     
     
         6 . The method of  claim 5 , wherein the set of at least nine microsatellite loci is co-amplified using at least one primer for each locus selected from the group consisting of: 
 SEQ ID NO: 1 and SEQ ID NO: 60 when the locus is BAT-25,    SEQ ID NO: 61 and SEQ ID NO: 62 when the locus is BAT-26,    SEQ ID NO: 65 and SEQ ID NO: 66 when the locus is MONO-15,    SEQ ID NO: 67 and SEQ ID NO: 68 when the locus is BAT-40,    SEQ ID NO: 63 and SEQ ID NO: 59 when the locus is D3S2432,    SEQ ID NO: 51 and SEQ ID NO: 52 when the locus is D7S1808,    SEQ ID NO: 53 and SEQ ID NO: 54 when the locus is D7S3070,    SEQ ID NO: 55 and SEQ ID NO: 64 when the locus is D7S3046, and    SEQ ID NO: 57 and SEQ ID NO: 58 when the locus is D10S1426.    
     
     
         7 . The method of  claim 1 , wherein the set of at least three microsatellite loci is co-amplified in step (c) using at least one oligonucleotide primer for each locus which is fluorescently labeled.  
     
     
         8 . 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.    
     
     
         9 . The method of  claim 8 , wherein the microsatellite instability results are used in prognostic tumor diagnosis.  
     
     
         10 . The method of  claim 8 , wherein the microsatellite instability results are used in the diagnosis of familial tumor predisposition.  
     
     
         11 . The method of  claim 8 , wherein the microsatellite instability results are used to detect cancerous tumors of the gastro-intestinal system and of the endometrium.  
     
     
         12 . The method of  claim 11  wherein the cancerous tumors are tumors from a colorectal cancer.  
     
     
         13 . A method of detecting microsatellite instability in genomic DNA, comprising the steps of: 
 a) providing primers for co-amplifying a set of at least three microsatellite loci of genomic DNA, comprising at least one mononucleotide repeat locus selected from the group consisting of BAT-25, BAT-26, BAT-40, MONO-11, and MONO-15, and at least two tetranucleotide repeat loci selected from the group consisting of FGA, D1S518, D1S547, D1S1677, D2S1790, D3S2432, D5S818, D5S2849, D6S1053, D7S3046, D7S1808, D7S3070, D8S1179, D9S2169, D10S1426, D10S2470, D12S391, D17S1294, D17S1299, and D18S51;    b) co-amplifying the set of at least three microsatellite loci from a first sample of genomic DNA originating from normal non-cancerous biological material from an individual and a from a second sample of genomic DNA originating from a second biological material from the individual, in separate multiplex amplification reactions, using at least one oligonucleotide primer for each of the microsatellite loci, thereby producing first amplified DNA fragments from the first sample and second amplified DNA fragments from the second sample; and    c) comparing the size of first amplified DNA fragments to the size of the second amplified DNA fragments to detect instability in any of the at least three microsatellite loci of the second genomic DNA.    
     
     
         14 . The method of  claim 13 , wherein the genomic DNA is human genomic DNA.  
     
     
         15 . The method of  claim 13 , wherein the set of at least three microsatellite loci is a set of at least five microsatellite loci, comprising two mononucleotide repeat loci selected from the group consisting of BAT-25, BAT-26, BAT-40, MONO-11, and MONO-15, and three tetranucleotide repeat loci selected from the group consisting of FGA, D1S518, D1S547, D1S1677, D2S1790, D3S2432, D5S5818, D5S2849, D6S1053, D7S3046, D7S1808, D7S3070, D8S1179, D9S2169, D10S1426,D10S2470,D12S391, D17S1294, D17S1299, and D18S51.  
     
     
         16 . The method of  claim 13 , wherein at least one of the primers provided in step (a) has a nucleic acid sequence selected from the group of primer sequences identified by: SEQ ID NO: 1-68.  
     
     
         17 . The method of  claim 13 , wherein the at least one primer for each locus provided in step (a) is fluorescently labeled.  
     
     
         18 . The method of  claim 13 , wherein the set of at least three microsatellite loci is a set of at least nine microsatellite loci, comprising: BAT-25, BAT-26, MONO-15, BAT-40, D3S2432, D7S3046, D7S3070, D7S1808, and D10S1426.  
     
     
         19 . The method of  claim 18 , wherein the set of at least nine microsatellite loci is co-amplified using at least one primer for each locus selected from the group consisting of: 
 SEQ ID NO: 1 and SEQ ID NO: 60 when the locus is BAT-25,    SEQ ID NO: 61 and SEQ ID NO: 62 when the locus is BAT-26,    SEQ ID NO: 65 and SEQ ID NO: 66 when the locus is MONO-15,    SEQ ID NO: 67 and SEQ ID NO: 68 when the locus is BAT-40,    SEQ ID NO: 63 and SEQ ID NO: 59 when the locus is D3S2432,    SEQ ID NO: 51 and SEQ ID NO: 52 when the locus is D7S1808,    SEQ ID NO: 53 and SEQ ID NO: 54 when the locus is D7S3070,    SEQ ID NO: 55 and SEQ ID NO: 64 when the locus is D7S3046, and    SEQ ID NO: 57 and SEQ ID NO: 58 when the locus is D10S1426.    
     
     
         20 . The method of  claim 13 , wherein the second sample of biological material is selected from the group consisting of: tumor tissue, disseminated cells, feces, blood cells, blood plasma, serum, lymph nodes, urine, and other bodily fluids.  
     
     
         21 . The method of  claim 13 , wherein the microsatellite instability results are used in prognostic tumor diagnosis.  
     
     
         22 . The method of  claim 13 , wherein the microsatellite instability results are used in the diagnosis of familial tumor predisposition.  
     
     
         23 . The method of  claim 13 , wherein the microsatellite instability results are used to detect cancerous tumors of the gastro-intestinal system and of the endometrium.  
     
     
         24 . The method of  claim 23 , wherein the cancerous tumors are tumors from a colorectal cancer.  
     
     
         25 . A kit for analyzing microsatellite loci of genomic DNA, comprising: 
 a single container with oligonucleotide primers for co-amplifying a set of at least three microsatellite loci of genomic DNA, the set comprising one mononucleotide repeat locus selected from the group consisting of BAT-25, BAT-26, BAT-40, MONO-11, and MONO-15, and at least two tetranucleotide repeat loci selected from the group consisting of FGA, D1S518, D1S547, D1S1677, D2S1790, D3S2432, D5S818, D5S2849, D6S1053, D7S3046, D7S1808, D7S3070, D8S1179, D9S2169, D10S1426, D10S2470, D12S391, D17S1294, D17S1299, and D18S51.    
     
     
         26 . The kit of  claim 25 , wherein the genomic DNA is human genomic DNA.  
     
     
         27 . The kit of  claim 25 , wherein the set of at least three microsatellite loci is a set of at least five microsatellite loci, comprising two mononucleotide repeat loci selected from the group consisting of BAT-25, BAT-26, BAT-40, MONO-11, and MONO-15, and three tetranucleotide repeat loci selected from the group consisting of FGA, D1S518, D1S547, D1S1677, D2S1790, D3S2432, D5S818, D5S2849, D6S1053, D7S3046, D7S1808, D7S3070, D8S1179, D9S2169, D10S1426, D10S2470, D12S391, D17S1294, D17S1299, and D18S51.  
     
     
         28 . The kit of  claim 25 , wherein the set of at least two microsatellite loci is a set of at least nine microsatellite loci, comprising: BAT-25, BAT-26, BAT-40, MONO-15, D3S2432, D7S1808, D7S3070, D7S3046, and D10S1426.  
     
     
         29 . The kit of  claim 28 , wherein at least one of the oligonucleotide primers is selected from the group consisting of: 
 SEQ ID NO: 1 and SEQ ID NO: 60 when the locus is BAT-25,    SEQ ID NO: 61 and SEQ ID NO: 62 when the locus is BAT-26,    SEQ ID NO: 65 and SEQ ID NO: 66 when the locus is MONO-15,    SEQ ID NO: 67 and SEQ ID NO: 68 when the locus is BAT-40,    SEQ ID NO: 63 and SEQ ID NO: 59 when the locus is D3S2432,    SEQ ID NO: 51 and SEQ ID NO: 52 when the locus is D7S1808,    SEQ ID NO: 53 and SEQ ID NO: 54 when the locus is D7S3070,    SEQ ID NO: 55 and SEQ ID NO: 64 when the locus is D7S3046, and    SEQ ID NO: 57 and SEQ ID NO: 58 when the locus is D10S1426.    
     
     
         30 . The kit of  claim 25 , wherein at least one oligonucleotide primer for each of the at least three microsatellite loci in the set is fluorescently labeled.  
     
     
         31 . The kit of  claim 25 , wherein the kit further comprises a thermostable polymerase.  
     
     
         32 . The kit of  claim 25 , wherein the kit further comprises first control DNA isolated from normal non-cancerous biological material, and second control DNA from biological material lacking mismatch repair genes.  
     
     
         33 . The kit of  claim 25 , wherein instability in the set of microsatellite loci co-amplified by the primers can be used in prognostic tumor diagnosis.  
     
     
         34 . The kit of  claim 25 , wherein instability in the set of microsatellite loci co-amplified by the primers can be used in the diagnosis of familial tumor predisposition.  
     
     
         35 . The kit of  claim 25 , wherein instability in the set of microsatellite loci co-amplified by the primers is an indication of tumors of the gastrointestinal system or of the endometrium.  
     
     
         36 . The kit of  claim 25 , wherein the microsatellite instability is an indication of colorectal cancer.  
     
     
         37 . A method of analyzing micro-satellite loci, comprising steps of: 
 a) providing primers for co-amplifying a set of at least nine microsatellite loci of human genomic DNA, comprising BAT-25, BAT-26, MONO-15, BAT-40, D3S2432, D7S3046, D7S3070, D7S1808, and D10S1426;    b) co-amplifying the set of at least three microsatellite 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.    
     
     
         38 . The method of  claim 37 , wherein the set of at least nine microsatellite loci is co-amplified using at least one primer for each locus selected from the group consisting of: 
 SEQ ID NO: 1 and SEQ ID NO: 60 when the locus is BAT-25,    SEQ ID NO: 61 and SEQ ID NO: 62 when the locus is BAT-26,    SEQ ID NO: 65 and SEQ ID NO: 66 when the locus is MONO-15,    SEQ ID NO: 67 and SEQ ID NO: 68 when the locus is BAT-40,    SEQ ID NO: 63 and SEQ ID NO: 59 when the locus is D3S2432,    SEQ ID NO: 51 and SEQ ID NO: 52 when the locus is D7S1808,    SEQ ID NO: 53 and SEQ ID NO: 54 when the locus is D7S3070,    SEQ ID NO: 55 and SEQ ID NO: 64 when the locus is D7S3046, and    SEQ ID NO: 57 and SEQ ID NO: 58 when the locus is D10S1426.    
     
     
         39 . A kit for analyzing microsatellite loci of genomic DNA, comprising: 
 a single container with oligonucleotide primers for co-amplifying a set of at least nine microsatellite loci of human genomic DNA, comprising BAT-25, BAT-26, MONO-15, BAT-40, D3S2432, D7S3046, D7S3070, D7S1808, and D10S1426.    
     
     
         40 . The kit of  claim 39 , wherein at least one of the oligonucleotide primers is selected from the group consisting of: 
 SEQ ID NO: 1 and SEQ ID NO: 60 when the locus is BAT-25,    SEQ ID NO: 61 and SEQ ID NO: 62 when the locus is BAT-26,    SEQ ID NO: 65 and SEQ ID NO: 66 when the locus is MONO-15,    SEQ ID NO: 67 and SEQ ID NO: 68 when the locus is BAT-40,    SEQ ID NO: 63 and SEQ ID NO: 59 when the locus is D3S2432,    SEQ ID NO: 51 and SEQ ID NO: 52 when the locus is D7S1808,    SEQ ID NO: 53 and SEQ ID NO: 54 when the locus is D7S3070,    SEQ ID NO: 55 and SEQ ID NO: 64 when the locus is D7S3046, and    SEQ ID NO: 57 and SEQ ID NO: 58 when the locus is D10S1426.    
     
     
         41 . A method of analyzing micro-satellite loci, comprising steps of: 
 a) providing primers for co-amplifying a set of at least three microsatellite loci of genomic DNA, comprising at least one mono-nucleotide repeat locus and at least two tetra-nucleotide repeat loci;    b) co-amplifying t he set of a t least three microsatellite 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.    
     
     
         42 . The method of  claim 41 , wherein the genomic DNA is human genomic DNA.  
     
     
         43 . The method of  claim 42 , wherein the at least two tetra-nucleotide repeat loci are selected from the group consisting of FGA, D1S518, D1S547, D1S1677, D2S1790, D3S2432, D5S818, D5S2849, D6S1053, D7S3046, D7S1801, D7S3070, D8S1179, D9S2169, D10S1426, D10S2470, D12S391, D17S1294, D17S1299, and D18S51.  
     
     
         44 . The method of  claim 42 , wherein the at least one mono-nucleotide repeat locus is selected from the group consisting of BAT-25, BAT-26, BAT-40, MONO-11, and MONO-15.  
     
     
         45 . The method of  claim 42 , wherein the set of at least three microsatellite loci is a set of at least five microsatellite loci, comprising: 
 at least two mono-nucleotide repeat loci selected from the group consisting of BAT-25, BAT-26, MONO-11, BAT-40, and MONO-15; and    at least three tetra-nucleotide repeat loci selected from the group consisting of FGA, D1S518, D1S547, D1S1677, D2S1790, D3S2432, D5S818, D5S2849, D6S1053, D7S3046, D7S1808, D7S3070, D8S1179, D9S2169, D10S1426, D10S2470, D12S391, D17S1294, D17S1299, and D18S51.    
     
     
         46 . The method of  claim 42 , wherein at least one of the primers provided in step (a) has a nucleic acid sequence selected from the group of primer sequences identified by: SEQ ID NO: 1-68.  
     
     
         47 . The method of  claim 42 , wherein the set of at least three microsatellite loci is a set of at least nine microsatellite loci, comprising BAT-25, BAT-26, MONO-15, D1S518, D3S2432, D7S1808, D7S3070, D7S3046, D10S1426.  
     
     
         48 . The method of  claim 47 , wherein the set of at least nine microsatellite loci is co-amplified using at least one primer for each locus selected from the group consisting of: 
 SEQ ID NO: 1 and SEQ ID NO: 60 when the locus is BAT-25,    SEQ ID NO: 61 and SEQ ID NO: 62 when the locus is BAT-26,    SEQ ID NO: 7 and SEQ ID NO: 8 when the locus is MONO-15,    SEQ ID NO: 49 and SEQ ID NO: 50 when the locus is D1S518,    SEQ ID NO: 17 and SEQ ID NO: 59 when the locus is D3S2432,    SEQ ID NO: 51 and SEQ ID NO: 52 when the locus is D7S1808,    SEQ ID NO: 53 and SEQ ID NO: 54 when the locus is D7S3070,    SEQ ID NO: 55 and SEQ ID NO: 56 when the locus is D7S3046, and    SEQ ID NO: 57 and SEQ ID NO: 58 when the locus is D10S1426.    
     
     
         49 . The method of  claim 42 , wherein the set of at least three microsatellite loci is a set of at least nine microsatellite loci, comprising BAT-25, BAT-26, MONO-15, BAT-40, D3S2432, D7S3046, D7S3070, D7S1808, and D10S1426.  
     
     
         50 . The method of  claim 49 , wherein the set of at least nine microsatellite loci is co-amplified using at least one primer for each locus selected from the group consisting of: 
 SEQ ID NO: 1 and SEQ ID NO: 60 when the locus is BAT-25,    SEQ ID NO: 61 and SEQ ID NO: 62 when the locus is BAT-26,    SEQ ID NO: 65 and SEQ ID NO: 66 when the locus is MONO-15,    SEQ ID NO: 67 and SEQ ID NO: 68 when the locus is BAT-40,    SEQ ID NO: 63 and SEQ ID NO: 59 when the locus is D3S2432,    SEQ ID NO: 51 and SEQ ID NO: 52 when the locus is D7S1808,    SEQ ID NO: 53 and SEQ ID NO: 54 when the locus is D7S3070,    SEQ ID NO: 55 and SEQ ID NO: 64 when the locus is D7S3046, and    SEQ ID NO: 57 and SEQ ID NO: 58 when the locus is D7S1426.    
     
     
         51 . The method of  claim 41 , wherein the set of at least three microsatellite loci is co-amplified in step (c) using at least one oligonucleotide primer for each locus which is fluorescently labeled.  
     
     
         52 . The method of  claim 41 , 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.    
     
     
         53 . The method of  claim 52 , wherein the microsatellite instability results are used in prognostic tumor diagnosis.  
     
     
         54 . The method of  claim 52 , wherein the microsatellite instability results are used in the diagnosis of familial tumor predisposition.  
     
     
         55 . The method of  claim 52 , wherein the microsatellite instability results are used to detect cancerous tumors of the gastro-intestinal system and of the endometrium.  
     
     
         56 . The method of  claim 55  wherein the cancerous tumors are tumors from a colorectal cancer.  
     
     
         57 . A kit for analyzing microsatellite loci of human genomic DNA, comprising: 
 a single container with oligonucleotide primers for co-amplifying a set of at least three microsatellite loci of human genomic DNA, the set comprising one mono-nucleotide repeat locus and two tetra-nucleotide repeat loci.    
     
     
         58 . The kit of  claim 57 , wherein the two tetra-nucleotide repeat loci are selected from the group consisting of FGA, D1S518, D1S547, D1S1677, D2S1790, D3S2432, D5S818, D5S2849, D6S1053, D7S3046, D7S1808, D7S3070, D8S1179, D9S2169, D10S1426, D10S2470, D12S391, D17S1294, D17S1299, and D18S51.  
     
     
         59 . The kit of  claim 57 , wherein the mono-nucleotide repeat locus is selected from the group consisting of BAT-25, BAT-26, BAT-40, MONO-11, and MONO-15.  
     
     
         60 . The kit of  claim 57 , wherein the set of at least three microsatellite loci is a set of at least five microsatellite loci, comprising two mono-nucleotide repeat loci selected from the group consisting of BAT-25, BAT-26, BAT-40, MONO-11, and MONO-15, and three tetra-nucleotide repeat loci selected from the group consisting of FGA, D1S518, D1S547, D1S1677, D2S1790, D3S2432, D5S818, D5S2849, D6S1053, D7S3046, D7S1808, D7S3070, D8S1179, D9S2169, D10S1426, D10S2470, D12S391, D17S1294, D17S1299, and D18S51.  
     
     
         61 . The kit of  claim 57 , wherein the set of at least two microsatellite loci is a set of at least nine microsatellite loci, comprising: BAT-25, BAT-26, MONO-15, D1S518, D3S2432, D7S1808, D7S3070, D7S3046, D10S1426.  
     
     
         62 . The kit of  claim 61 , wherein at least one of the oligonucleotide primers is selected from the group consisting of: 
 SEQ ID NO: 1 and SEQ ID NO: 60 when the locus is BAT-25,    SEQ ID NO: 61 and SEQ ID NO: 62 when the locus is BAT-26,    SEQ ID NO: 7 and SEQ ID NO: 8 when the locus is MONO-15,    SEQ ID NO: 49 and SEQ ID NO: 50 when the locus is D1S518,    SEQ ID NO: 17 and SEQ ID NO: 59 when the locus is D3S2432,    SEQ ID NO: 51 and SEQ ID NO: 52 when the locus is D7S1808,    SEQ ID NO: 53 and SEQ ID NO: 54 when the locus is D7S3070,    SEQ ID NO: 55 and SEQ ID NO: 56 when the locus is D7S3046, and    SEQ ID NO: 57 and SEQ ID NO: 58 when the locus is D10S1426.    
     
     
         63 . The kit of  claim 57 , wherein the set of at least two microsatellite loci is a set of at least nine microsatellite loci, comprising: BAT-25, BAT-26, MONO-15, BAT-40, D3S2432, D7S3046, D7S3070, D7S1808, and D 10S1426.  
     
     
         64 . The kit of  claim 63 , wherein at least one of the oligonucleotide primers is selected from the group consisting of: 
 SEQ ID NO: 1 and SEQ ID NO: 60 when the locus is BAT-25,    SEQ ID NO: 61 and SEQ ID NO: 62 when the locus is BAT-26,    SEQ ID NO: 65 and SEQ ID NO: 66 when the locus is MONO-15,    SEQ ID NO: 67 and SEQ ID NO: 68 when the locus is BAT-40,    SEQ ID NO: 63 and SEQ ID NO: 59 when the locus is D3S2432,    SEQ ID NO: 51 and SEQ ID NO: 52 when the locus is D7S1808,    SEQ ID NO: 53 and SEQ ID NO: 54 when the locus is D7S3070,    SEQ ID NO: 55 and SEQ ID NO: 64 when the locus is D7S3046, and    SEQ ID NO: 57 and SEQ ID NO: 58 when the locus is D10S1426.    
     
     
         65 . The kit of  claim 57 , wherein at least one oligonucleotide primer for each of the at least three microsatellite loci in the set is fluorescently labeled.  
     
     
         66 . The kit of  claim 57 , wherein the kit further comprises a thermostable polymerase.  
     
     
         67 . The kit of  claim 57 , wherein the kit further comprises first control DNA isolated from normal non-cancerous biological material, and second control DNA from biological material lacking mismatch repair genes.  
     
     
         68 . The kit of  claim 57 , wherein instability in the set of microsatellite loci co-amplified by the primers can be used in prognostic tumor diagnosis.  
     
     
         69 . The kit of  claim 57 , wherein instability in the set of microsatellite loci co-amplified by the primers can be used in the diagnosis of familial tumor predisposition.  
     
     
         70 . The kit of  claim 57 , wherein instability in the set of microsatellite loci co-amplified by the primers is an indication of tumors of the gastrointestinal system or of the endometrium.  
     
     
         71 . The kit of claim  70 , wherein the microsatellite instability is an indication of colorectal cancer.