HLA-DR8 Subtyping by Polymerase Chain Reaction (PCR) - DNA Conformation Polymorphism (DCP) Analysis : a simple and practical genotyping method
Hiroshima Journal of Medical Sciences 45 巻 3 号
85-92 頁
1996-09 発行
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ファイル情報(添付) | |
タイトル ( eng ) |
HLA-DR8 Subtyping by Polymerase Chain Reaction (PCR) - DNA Conformation Polymorphism (DCP) Analysis : a simple and practical genotyping method
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作成者 |
Fukuda Yasuhiko
Kimura Akinori
Hoshino Shuji
Shintaku Sadanori
Sakaguchi Takemasa
Asahara Toshimasa
Sakaki Miyoko
Sumimoto Ryo
Furukawa Masahiro
Ohdan Hideki
Yoshida Tetsuya
Dohi Kiyohiko
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収録物名 |
Hiroshima Journal of Medical Sciences
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巻 | 45 |
号 | 3 |
開始ページ | 85 |
終了ページ | 92 |
収録物識別子 |
[PISSN] 0018-2052
[EISSN] 2433-7668
[NCID] AA00664312
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抄録 |
Two hundred Japanese panels were serologically typed for human leukocyte antigen (HLA) - DR to assign 65 HLA-DRS haplotypes, which were then subdivided into two genotypes, i.e., DRB1 *0802 and DRB1 *0803, by a polymerase chain reaction (PCR) - based, simple, and practical method. The panels possessing DR8 specificity were firstly subjected to PCR with a couple of primers specifically to amplify their DR52 associated group - DRB1 genes. PCR products were then denatured in the presence of formamide, electrophoresed in a non-denaturing polyacrylamide gel, and visualized by silver staining. The same DRB1 products of these samples were also mixed with the DRB1 *1302, and simultaneously analyzed by the same procedure. Electrophoretic mobilities of the samples were compared with those of the typing standards to genotype their DRS-DRB1 alleles by using the characteristic polymorphism in the single-stranded DNAs and the heteroduplexes. This method, designated PCR - DNA conformation polymorphism (DCP) analysis, allowed for genotyping of the DR8-DRB1 alleles without using sequence - specific oligonucleotide probes (SSOP) or restriction endonucleases. The entire process after PCR was completed within a few hours. The tested panels were also genotyped for DRB1 gene by the PCR-SSOP method for comparison with results obtained by the PCR-DCP method. Satisfactory coincidence was achieved and it represented how accurately the new system genotyped DRB1 *0802 and DRB1 *0803. PCR-DCP analysis was thus shown to be practical and useful for subtyping of serologically defined DR8 specificities.
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著者キーワード |
PCR (polymerase chain reaction)
Subcloning, HLA (human leukocyte antigen)
SSO (sequence-specific oligonucleotide) typing
Sanger's dideoxy sequencing method
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内容記述 |
This work was supported in part by grants-in-aid from the Ministry of Education, Culture and Science, Japan, and research grants from the Ministry of Health and Welfare, Japan.
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NDC分類 |
医学 [ 490 ]
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言語 |
英語
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資源タイプ | 紀要論文 |
出版者 |
Hiroshima University Medical Press
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発行日 | 1996-09 |
出版タイプ | Version of Record(出版社版。早期公開を含む) |
アクセス権 | オープンアクセス |
収録物識別子 |
[ISSN] 0018-2052
[NCID] AA00664312
|