Method for detecting cystic fibrosis

Abstract

The present invention relates to methods for amplifying various regions of the cystic fibrosis transmembrane regulator (CFTR) gene. Methods are provided for amplifying one or all 27 exons of the CFTR gene and a portion of the CFTR promoter region in a single tube. The method can identify the presence or absence of CF deletions or insertions in a sample and assist in the diagnosis of a genetic predisposition to cystic fibrosis.

1 .- 22 . (canceled) 
     
     
         23 . A kit comprising oligonucleotide primer pairs suitable for amplifying multiple target segments of a CFTR gene, wherein at least one primer of each oligonucleotide primer pair is detectably labeled, and wherein the kit comprises at least one primer selected from the group consisting of SEQ ID NOs: 1-70. 
     
     
         24 . The kit of  claim 23 , wherein the kit comprises oligonucleotide primer pairs suitable for amplifying at least seven different target segments of the human CFTR gene. 
     
     
         25 . The kit of  claim 23 , wherein the kit comprises oligonucleotide primer pairs suitable for amplifying a target segment from each of exon 22, exon 23, and exon 24 of the human CFTR gene. 
     
     
         26 . The kit of  claim 25 , wherein:
 (a) the oligonucleotide primer pair for amplification of a target segment of exon 22 of the CFTR gene consists of a forward primer comprising the sequence of SEQ ID NO: 51 and/or a reverse primer comprising the sequence of SEQ ID NO: 52;   (b) the oligonucleotide primer pair for amplification a target segment of exon 23 of the CFTR gene consists of a forward primer comprising the sequence of SEQ ID NO: 53 and/or a reverse primer comprising the sequence of SEQ ID NO: 54; and/or   (c) the oligonucleotide primer pair for amplification a target segment of exon 24 of the CFTR gene consists of a forward primer comprising the sequence of SEQ ID NO: 55 and a reverse primer comprising the sequence of SEQ ID NO: 56.   
     
     
         27 . The kit of  claim 23 , wherein at least one primer comprises a sequence selected from the group consisting of SEQ ID NOs: 51, 52, 53, 54, 55 and 56. 
     
     
         28 . The kit of  claim 23 , wherein the kit comprises the primers of SEQ ID NOs: 51, 52, 53, 54, 55 and 56. 
     
     
         29 . The kit of  claim 23 , wherein the kit comprises oligonucleotide primer pairs suitable for amplifying a target segment from each of the promoter, exon 1, exon 2, exon 22, exon 23, and exon 24 of the human CFTR gene. 
     
     
         30 . The kit of  claim 29 , wherein:
 (a) the oligonucleotide primer pair for amplification of a target segment of the promoter of the CFTR gene consists of a forward primer comprising the sequence of SEQ ID NO: 1 and/or a reverse primer comprising the sequence of SEQ ID NO: 2;   (b) the oligonucleotide primer pair for amplification of a target segment of exon 1 of the CFTR gene consists of a forward primer comprising the sequence of SEQ ID NO: 3 and/or a reverse primer comprising the sequence of SEQ ID NO: 4.   (d) the oligonucleotide primer pair for amplification of a target segment of exon 22 of the CFTR gene consists of a forward primer comprising the sequence of SEQ ID NO: 51 and/or a reverse primer comprising the sequence of SEQ ID NO: 52;   (d) the oligonucleotide primer pair for amplification a target segment of exon 23 of the CFTR gene consists of a forward primer comprising the sequence of SEQ ID NO: 53 and/or a reverse primer comprising the sequence of SEQ ID NO: 54; and/or   (e) the oligonucleotide primer pair for amplification a target segment of exon 24 of the CFTR gene consists of a forward primer comprising the sequence of SEQ ID NO: 55 and a reverse primer comprising the sequence of SEQ ID NO: 56.   
     
     
         31 . The method according to  claim 23 , wherein the kit comprises oligonucleotide primer pairs suitable for amplifying at least 28 target segments of the CFTR gene. 
     
     
         32 . The method according to  claim 31 , wherein the at least 28 target segments together comprise sequence from all or a portion of 20 different exons of the CFTR gene. 
     
     
         33 . The kit of  claim 23 , wherein the label is a fluorescent label.