Patent EP-2447378-A1 (EP)
Inventor: IGUCHI AKI
IPC: C12Q1/68
Filed: 2011-10-28
Abstract: A polymorphism-detecting probe, an amplification primer and the use thereof are provided to enable simple and highly reliable determination of different polymorphisms in an EGFR gene. 
A fluorescence-labeled oligonucleotide selected from at least one of P1, P3, P5 - P7, and P15 - P18 below is used as the polymorphism-detecting probe. 
(P1) An oligonucleotide comprising or consisting of a complementary sequence to a nucleotide sequence having a length of 11 - 50 bases comprising base positions 251 - 261 of the base sequence of SEQ ID NO: 1 or a sequence having homology with said complementary sequence, and in which the base corresponding to base number 261 is adenine or cytosine, the base corresponding to base number 251 is cytosine, and the cytosine is fluorescence-labeled; 
(P3) An oligonucleotide comprising or consisting of a complementary sequence to a nucleotide sequence having a length of 5 - 50 bases comprising base positions 257 - 261 of the base sequence of SEQ ID NO: 1 or a sequence having homology with said complementary sequence, and in which the base corresponding to base number 261 is adenine or cytosine, the base corresponding to base number 257 is cytosine, and the cytosine is fluorescence-labeled; 
(P5) An oligonucleotide comprising or consisting of a sequence having homology with a nucleotide sequence having a length of 9 - 50 bases comprising base positions 104 - 112 of the base sequence of SEQ ID NO: 2, and in which the base that is homologous to base number 112 is thymine, the base that is homologous to base number 104 is cytosine, and the cytosine is fluorescence-labeled; 
(P6) An oligonucleotide comprising or consisting of a sequence having homology with a nucleotide sequence having a length of 16 - 50 bases comprising base positions 104 - 119 of the base sequence of SEQ ID NO: 2, and in which the base at position 119 is substituted by a base other than guanine, and in which the base that is homologous to base number 104 is cytosine, and the cytosine is fluorescence-labeled; 
(P7) An oligonucleotide comprising or consisting of a sequence having homology with a nucleotide sequence having a length of 10 - 50 bases comprising base positions 136 - 145 of the base sequence of SEQ ID NO: 3, and in which the base that is homologous to base number 145 is cytosine, and the cytosine is fluorescence-labeled; 
(P 15) An oligonucleotide comprising or consisting of a complementary sequence to a nucleotide sequence having a length of 6 - 50 bases comprising base positions 259 - 264 of the base sequence of SEQ ID NO: 1 or a sequence having homology with said complementary sequence, and in which the base corresponding to base number 261 is adenine or cytosine, and cytosine positioned at the 3' side relative to the base is fluorescence-labeled; 
(P16) An oligonucleotide comprising or consisting of a complementary sequence to a nucleotide sequence having a length of 5 - 50 bases comprising base positions 258 - 262 of the base sequence of SEQ ID NO: 1 or a sequence having homology with said complementary sequence, and in which the base corresponding to base number 261 is adenine or cytosine, and cytosine positioned at the 5' side relative to the base is fluorescence-labeled; 
(P17) An oligonucleotide comprising or consisting of a sequence having homology with a nucleotide sequence having a length of 16 - 50 bases comprising base positions 249 - 264 of the base sequence of SEQ ID NO: 1, and in which the base that is homologous to base number 261 is thymine or guanine, and cytosine positioned at the 5' side relative to the base is fluorescence-labeled; and 
(P18) An oligonucleotide comprising or consisting of a sequence having homology with a nucleotide sequence having a length of 8 - 50 bases comprising base positions 257 - 264 of the base sequence of SEQ ID NO: 1, and in which the base that is homologous to base number 261 is thymine or guanine, and cytosine positioned at the 3' side relative to the base is fluorescence-labeled.