Nisshin Seifun Group Inc.

Nippon Flour Mills Co., Ltd

Shinjiro Imai

Keiko Tanaka

Youichi Kurimoto

Hiroyuki Haraguchi

Kazumi Kitta

Yasuyuki Matsuoka

Junichi Mano

Shinichiro Arami

Satoshi Furui

Megumi Sato

Provided is a method of detecting or quantifying a wheat species-specific DNA in a test sample by polymerase chain reaction. The method comprises a step of amplifying a nucleic acid molecule having a partial sequence of a nucleotide sequence identified as SEQ ID NO: 1 using a nucleic acid molecule in the test sample or a nucleic acid molecule extracted from the test sample as the template and using a primer pair capable of amplifying the partial sequence and a step of detecting or quantifying the amplified nucleic acid molecule.

Method for detecting and quantifying wheat endogenous gene

Provided is a method of detecting or quantifying a wheat species-specific DNA in a test sample by polymerase chain reaction. The method comprises a step of amplifying a nucleic acid molecule having at partial sequence of a nucleotide sequence identified as SEQ ID NO: 1 using a nucleic acid molecule in the test sample or a nucleic acid molecule extracted from the test sample as the template and using a primer pair capable of amplifying the partial sequence and a step of detecting or quantifying the amplified nucleic acid molecule.

Disclosed are: a method for detecting common wheat among from wheat varieties contained in a sample of interest such as a food raw material or a processed food specifically, with high sensitivity, and in a qualitative and/or quantitative manner; a method for discriminating between common wheat and a wheat variety other than common wheat (e.g., durum wheat) contained in a food raw material or a processed food and detecting the common wheat in a qualitative and/or quantitative manner; and a primer set, a nucleic acid probe, and a detection kit, each of which can be used in the methods employing a PCR method. Specifically disclosed are: a method for detecting the occurrence of common wheat in a sample of interest, which comprises carrying out a PCR method using a nucleic acid extracted from the sample as a template and using a primer comprising the nucleotide sequence represented by SEQ ID NO:5 and a primer comprising the nucleotide sequence represented by SEQ ID NO:6 and detecting the occurrence of a PCR amplification product; and a method for detecting the occurrence of common wheat in a sample of interest, which comprises carrying out a quantitative PCR method using a nucleic acid extracted from the sample as a template and using a primer comprising the nucleotide sequence represented by SEQ ID NO:5, a primer comprising the nucleotide sequence represented by SEQ ID NO:6 and a nucleic acid probe comprising the nucleotide sequence represented by SEQ ID NO:11 and detecting the occurrence of common wheat qualitatively and/or quantitatively.

Method for qualitative and quantitative detection of common wheat

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 by 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 method of detecting or quantitating endogenous wheat DNA in a test sample by the polymerase chain reaction, the method comprising: a step of using a nucleic acid molecule in the test sample or a nucleic acid molecule extracted from the test sample as a template to amplify the nucleic acid molecule of a region consisting of the nucleotide sequence identified as SEQ ID NO: 2 or a partial sequence thereof with a primer pair capable of amplifying that region; and a step of detecting or quantitating the amplified nucleic acid molecule.

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Saitama, Japan

Shinjiro Imai