PMID- 9271827
OWN - NLM
STAT- MEDLINE
DCOM- 19971028
LR  - 20190826
IS  - 0001-2815 (Print)
IS  - 0001-2815 (Linking)
VI  - 50
IP  - 2
DP  - 1997 Aug
TI  - High-resolution HLA-DRB typing using denaturing gradient gel electrophoresis and 
      direct sequencing.
PG  - 170-7
AB  - High-resolution HLA-DRB typing is required for bone marrow transplantation 
      between unrelated donors and recipients and also for identification of novel 
      HLA-DRB alleles. Here we describe a method for the unambiguous identification of 
      HLA-DRB alleles using the polymerase chain reaction (PCR), denaturing gradient 
      gel electrophoresis (DGGE) and direct sequencing. The highly variable second exon 
      of all HLA-DRB1, -DRB3, -DRB4, -DRB5, -DRB6 and -DRB7 alleles was amplified using 
      a single pair of generic DRB-specific primers and alleles were separated by DGGE. 
      DNA was then reamplified from plugs removed from the gel and the sequences of 
      these alleles were determined using fluorescent-based sequencing and 
      allele-assignment software. The validity of this typing procedure was confirmed 
      by identification of HLA-DRB alleles for 17 individuals previously characterized 
      by PCR-SSP and/or cloning and sequencing techniques. We identified 34 different 
      HLA-DRB alleles in these 17 unrelated individuals. Importantly, our analysis 
      revealed HLA-DRB1 alleles which had not been identified using the PCR-SSP typing 
      technique. Additionally, alleles from the HLA-DRB3, -DRB4 and -DRB5 loci were 
      identified. Whereas traditional HLA-DRB typing methods provide limited 
      information or require the use of multiple oligonucleotide primers or probes, our 
      technique provides a reliable, specific and relatively rapid way of identifying 
      all HLA-DRB alleles for high-resolution tissue typing.
FAU - Knapp, L A
AU  - Knapp LA
AD  - Wisconsin Regional Primate Research Center, University of Wisconsin, Madison, 
      USA.
FAU - Lehmann, E
AU  - Lehmann E
FAU - Hennes, L
AU  - Hennes L
FAU - Eberle, M E
AU  - Eberle ME
FAU - Watkins, D I
AU  - Watkins DI
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Tissue Antigens
JT  - Tissue antigens
JID - 0331072
RN  - 0 (DNA Primers)
RN  - 0 (HLA-DR Antigens)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Alleles
MH  - DNA/analysis
MH  - DNA Primers/chemistry
MH  - Electrophoresis, Polyacrylamide Gel/*methods
MH  - Exons
MH  - HLA-DR Antigens/analysis/*classification
MH  - Histocompatibility Testing/methods
MH  - Humans
MH  - Nucleic Acid Denaturation
MH  - Polymerase Chain Reaction/methods
MH  - Sequence Analysis, DNA/*methods
EDAT- 1997/08/01 00:00
MHDA- 1997/08/01 00:01
CRDT- 1997/08/01 00:00
PHST- 1997/08/01 00:00 [pubmed]
PHST- 1997/08/01 00:01 [medline]
PHST- 1997/08/01 00:00 [entrez]
AID - 10.1111/j.1399-0039.1997.tb02856.x [doi]
PST - ppublish
SO  - Tissue Antigens. 1997 Aug;50(2):170-7. doi: 10.1111/j.1399-0039.1997.tb02856.x.