Method for determining HLA-A*24 group

A technology of HLA-A and determination method, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, recombinant DNA technology, etc., can solve the problems of labor-consuming, differences in HLA type distribution, etc., and achieve the effect of simple and accurate accuracy

Inactive Publication Date: 2014-09-03
TOPPAN PRINTING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

By performing multi-stage assays like this method, it is expected to eliminate ambiguity and perform detailed typing of HLA-A types, but there is a problem that it takes a lot of labor
[0006] In addition, the distribution of HLA types is different according to race

Method used

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  • Method for determining HLA-A*24 group
  • Method for determining HLA-A*24 group
  • Method for determining HLA-A*24 group

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] The HLA-A type is the same type of [A*24:02 / A*24:02], the heterotype of [A*24:02 / A*24:08], [A*24:20 / A*32: 01] and [A*32:01 / A*24:04] isotypes of subjects with genomic DNA extracted from the biological samples as a template, using the primer-1 and primer-2 shown in Table 8 to implement PCR, and PCR performed using primer-3 and primer-4, sequence analysis was performed on each PCR product.

[0092] Table 11 shows the results of the sequence analysis and the judgment results based on the obtained typing results. In the table, "+" in the column of "judgment" indicates the determination that the genomic DNA contains HLA-A*24:02, and "-" indicates the determination that the genomic DNA does not contain HLA-A*24:02, respectively. As a result, [A*24:02 / A*24:02] homotype, [A*24:02 / A*24:08] heterotype and [A*32:01 / A*24:04] heterotype are all was accurately judged. On the other hand, the outlier of [A*24:20 / A*32:01] is a false positive, as speculated.

[0093] Table 11

[0094...

Embodiment 2

[0096]Except using Primer-5 shown in Table 9 instead of Primer-4, the same operation was performed as in Example 1, so that the HLA-A type was [A*24:02 / A*24:02] isotype, [A *24:02 / A*24:08], [A*24:20 / A*32:01] and [A*32:01 / A*24:04] subjects Genomic DNA extracted from a biological sample is used as a template for PCR. As a result, by PCR using primer-3 and primer-5, PCR products were obtained even when any genomic DNA was used as a template. Therefore, the genomic DNA typing of all subjects is: the 437th base is C, the 441st base is T, the 443rd base is G, the 444th base is C, and the 447th base is C. The base is T, and the 449th base is C. When sequence analysis was performed on each PCR product, as shown in Table 12, the same determination results as in Example 8 were obtained. However, when the heterotyped genomic DNA from [A*32:01 / A*24:04] was used, the base (b) in Example 1 was typed into C and G, but in this embodiment The base in (b) in the example is only typed as C. ...

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Abstract

The present invention provides a method for determining the typing of an HLA-A*24 group with less ambiguity in a simple manner. The method according to the present invention is: a method for determining an HLA-A*24 group, characterized by determining the HLA-A*24 group on the basis of the results of the typing of (a) a 211th nucleotide and (b) a 395th nucleotide in the genome DNA of a subject, wherein A (adenine) in the initiation codon for HLA gene is defined as the 1st nucleotide; or the aforementioned method for determining an HLA-A*24 group, characterized by determining the HLA-A*24 group on the basis of the results of the typing of (c) at least one nucleotide selected from the group consisting of a 437th nucleotide, a 441st nucleotide, a 443rd nucleotide, a 444th nucleotide, a 447th nucleotide and a 449th nucleotide in addition to the results of the typing of the above-mentioned nucleotides (a) and (b).

Description

technical field [0001] The present invention relates to a method for judging whether the HLA type is HLA-A*24 group. [0002] This application claims priority based on Japanese Patent Application No. 2011-289336 filed in Japan on December 28, 2011, the content of which is hereby incorporated by reference. Background technique [0003] HLA (Human Leukocyte Antigen, Human Leukocyte Antigen) is a gene product of a gene cluster encoded in the MHC (Major Histocompatibility Complex: Major Histocompatibility Complex) region located on the short arm of the sixth chromosome. HLA has a very rich diversity, especially in clinical practice operations such as hematopoietic stem cell transplantation and blood transfusion, it is very important to improve the goodness of fit of HLA. Therefore, accurate HLA typing with as little labor as possible is required in clinical practice. [0004] HLA typing methods can be roughly divided into serological typing methods and DNA typing methods. Com...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12Q1/68
CPCC12Q1/6881C12Q2600/156
Inventor 牧野洋一山根明男东友彦中山雅人
Owner TOPPAN PRINTING CO LTD
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