Method of analyzing change in primary structure of nucleic acid

Abstract

It is intended to provide a method of analyzing a change in the primary structure of a nucleic acid which is excellent in quantification properties, achieves a high sensitivity and a high reproducibility and can be quickly conducted at a low cost. Namely, a method of analyzing a change in the primary structure of a nucleic acid characterized by comprising: the step of obtaining nucleic acids wherein a first ligand and a second ligand differing from each other in nucleic acid type are bound to the standard sequence in a standard nucleic acid and a target sequence in a subject nucleic acid to be analyzed; the step of specifically binding the first ligand to a receptor held on a support to thereby immobilize the standard sequence and the target sequence to the support; the step of specifically binding a labeled receptor to the second ligand in the nucleic acids thus immobilized; the step of detecting the label to thereby detect the standard sequence and the target sequence; the step of determining the ratio of the standard sequence in the standard nucleic acid and the target nucleic acid to be analyzed, thus calculating a coefficient for correcting the detection data of the target sequence followed by the correction; and the step of confirming an increase or a decrease in the amount of the target sequence in the subject nucleic acid to be analyzed compared with the target sequence in the standard nucleic acid.

Description

technical field [0001] The present invention relates to a method for analyzing changes in the primary structure of nucleic acids suitable for determining nucleic acid mutations. Background technique [0002] As the base sequence of the human genome is deciphered, positional information of genes on chromosomes based on the deciphered information becomes clear. In addition, based on this positional information, studies are being conducted toward practical diagnosis of various chromosomal structural changes that cause cancer, that is, primary structural changes of nucleic acids. In order to analyze changes in the primary structure of nucleic acids in a network, there is the array CGH method, which uses a microarray with hundreds to thousands, sometimes tens of thousands, of cDNA clones or BAC clones mapped on chromosomes, Nucleic acid is extracted, and the entire nucleic acid is amplified or not amplified, fluorescently labeled, and hybridized to analyze loss of heterozygotes ...

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Problems solved by technology

[0010] However, the analysis by the array CGH method described in Patent Document 1 and Non-Patent Document 1 has the problem that it is not practical as a method used in the clinical field because it takes time and cost to fabricate the array, and the experimental operation is also complicated.

In addition, since the entire nucleic acid extracted from cells is labeled and hybridized, noise may occur due to cross-hybridization, etc., and there is a problem that it is difficult to quantitatively obtain accurate data.

[0011] In addition, the analysis method using polymorphic markers and electrophoresis may be homozygous depending on the genotype of the patient, and qualitative analysis cannot be performed. In addition, the experimental operation is complicated and time-consuming, and it is difficult to accurately quantify the target gene. The problem that the method used in

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