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ID 31305
JaLCDOI
フルテキストURL
著者
Inoue, Seiichi Okayama University
Yamamoto, Yuji Okayama University
Okamoto, Osamu Okayama Univeristy
Murakami, Hiroki Okayama University
Miyaishi, Satoru Okayama University
Isizu, Hideo Osaka University
抄録

A sensitive method of HLA-DRB1 typing was devised using a semi-nested polymerase chain reaction (PCR) followed by a restriction fragment length polymorphism (RFLP) analysis (semi-nested PCR-RFLP method). The first-round amplification (30 cycles) of the semi-nested PCR was performed using DRB generic primer pairs and the second round of PCRs (20 cycles) were performed using DRB1 group-specific primers. The products of the second round PCRs were digested with restriction endonucleases for the typing of HLA-DRB1 alleles. By this method, HLA-DRB1 typing was possible from 10 pg of genomic DNA extracted from lymphocytes and from 0.5 microliter of 1,000 times diluted blood without DNA extraction. HLA-DRB1 alleles could be typed from a 2-mm long bloodstained cotton thread prepared from 10 times diluted blood and from a 2-mm thread of whole blood bloodstains stored at room temperature for 2 years. From the mixture of blood of two individuals with different genotypes, DRB1 alleles of the minor component were detected down to 1/1,000 of the major component. This semi-nested PCR-RFLP method is useful for HLA-DRB1 typing from extremely small amounts of DNA and from mixed samples.

キーワード
polymorphism
HLA-DRB1
polymerase chain reaction
dsmi-nested PCR
restricton fragment length polymotphism
Amo Type
Article
発行日
1998-12
出版物タイトル
Acta Medica Okayama
52巻
6号
出版者
Okayama University Medical School
開始ページ
289
終了ページ
296
ISSN
0386-300X
NCID
AA00508441
資料タイプ
学術雑誌論文
言語
English
論文のバージョン
publisher
査読
有り
PubMed ID
Web of Sience KeyUT