PMID- 8916570
OWN - NLM
STAT- MEDLINE
DCOM- 19970109
LR  - 20111117
IS  - 0018-2052 (Print)
IS  - 0018-2052 (Linking)
VI  - 45
IP  - 3
DP  - 1996 Sep
TI  - HLA-DR8 subtyping by polymerase chain reaction (PCR)-DNA conformation 
      polymorphism (DCP) analysis: a simple and practical genotyping method.
PG  - 85-92
AB  - Two hundred Japanese panels were serologically typed for human leukocyte antigen 
      (HLA) - DR to assign 65 HLA-DR8 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 DR8--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.
FAU - Fukuda, Y
AU  - Fukuda Y
AD  - 2nd Department of Surgery, Hiroshima University School of Medicine, Japan.
FAU - Kimura, A
AU  - Kimura A
FAU - Hoshino, S
AU  - Hoshino S
FAU - Shintaku, S
AU  - Shintaku S
FAU - Sakaguchi, T
AU  - Sakaguchi T
FAU - Asahara, T
AU  - Asahara T
FAU - Sakaki, M
AU  - Sakaki M
FAU - Sumimoto, R
AU  - Sumimoto R
FAU - Tashiro, H
AU  - Tashiro H
FAU - Furukawa, M
AU  - Furukawa M
FAU - Ohdan, H
AU  - Ohdan H
FAU - Yoshida, T
AU  - Yoshida T
FAU - Dohi, K
AU  - Dohi K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Japan
TA  - Hiroshima J Med Sci
JT  - Hiroshima journal of medical sciences
JID - 0421060
RN  - 0 (HLA-DR Antigens)
RN  - 0 (HLA-DR Serological Subtypes)
RN  - 0 (HLA-DR8 antigen)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Base Sequence
MH  - Cell Line
MH  - DNA/genetics
MH  - Evaluation Studies as Topic
MH  - Genotype
MH  - HLA-DR Antigens/*classification/*genetics
MH  - HLA-DR Serological Subtypes
MH  - Histocompatibility Testing/methods
MH  - Humans
MH  - Polymerase Chain Reaction/methods
MH  - *Polymorphism, Single-Stranded Conformational
EDAT- 1996/09/01 00:00
MHDA- 1996/09/01 00:01
CRDT- 1996/09/01 00:00
PHST- 1996/09/01 00:00 [pubmed]
PHST- 1996/09/01 00:01 [medline]
PHST- 1996/09/01 00:00 [entrez]
PST - ppublish
SO  - Hiroshima J Med Sci. 1996 Sep;45(3):85-92.