Method of detecting or quantitating endogenous wheat dna and method of determining contamination rate of genetically modified wheat in test sample

Abstract

An object of the present invention is to discover an endogenous wheat sequence satisfying the conditions of: a) it is universally present in varieties of wheat, b) the amount present (detected amount) is not affected depending on the wheat variety, c) even if other grains are present, only wheat can be detected without cross-reactivity, and d) it is amplified quantitatively by the PCR reaction. A further object of the present invention is to provide a method of accurately detecting and quantitating endogenous wheat DNA in a test sample by the polymerase chain reaction. The present invention provides a kit for detecting or quantitating an endogenous wheat DNA sequence in a test sample by the polymerase chain reaction, the kit comprising at least one primer pair capable of amplifying the endogenous wheat DNA sequence.

1 .- 13 . (canceled) 
     
     
         14 . A kit for detecting or quantitating an endogenous wheat DNA sequence in a test sample by the polymerase chain reaction, comprising at least one primer pair selected from
 (i) a primer pair consisting of a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 3 and a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 4;   (ii) a primer pair consisting of a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 5 and a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 4;   (iii) a primer pair consisting of a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 3 and a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 6;   (iv) a primer pair consisting of a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 5 and a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 6;   (v) a primer pair consisting of a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 3 and a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 7;   (vi) a primer pair consisting of a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 3 and a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 8;   (vii) a primer pair consisting of a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 5 and a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 8;   (viii) a primer pair consisting of a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 5 and a nucleic acid molecule comprising the nucleotide sequence identified as SEQ ID NO: 7; and   (ix) a primer pair consisting of a pair of nucleic acid molecules, wherein one nucleic acid molecule in the primer pair in (ix) comprises a nucleotide sequence in common with at least 80% continuous nucleotide sequence of one nucleic acid molecule in one of the primer pairs in (i) to (viii) above, and the other nucleic acid molecule in the primer pair in (ix) comprises a nucleotide sequence in common with at least 80% continuous nucleotide sequence of the other nucleic acid molecule in the same primer pair in (i) to (viii) above.