PMID- 7810876
OWN - NLM
STAT- MEDLINE
DCOM- 19950130
LR  - 20071006
IS  - 0003-2697 (Print)
IS  - 0003-2697 (Linking)
VI  - 221
IP  - 2
DP  - 1994 Sep
TI  - Homogeneous fluorescence detection method for human leukocyte antigen-DR typing 
      following polymerase chain reaction amplification with sequence-specific primer.
PG  - 340-7
AB  - A fluorescent homogeneous method for the detection of sequence-specific 
      amplification of human leukocyte antigen (HLA) alleles has been developed. In 
      this approach, polymerase chain reaction sequence-specific primers (PCR-SSP) are 
      used to amplify DRB1, DRB3, and DRB4 alleles. Lambda exonuclease and Exonuclease 
      I are added to reduce background by digesting template DNA, partial primer dimer, 
      and primer. PCR amplicons are then detected following the addition of a 
      fluorescent dye (thiazole yellow dimer) which binds to double-stranded DNA. No 
      transfer or wash steps are required. Thus, the risk of sample contamination, 
      which is a major source of inaccuracy for DNA amplification methods, is greatly 
      reduced. This approach is also faster and more easily automated than the standard 
      approach using gel electrophoresis and ethidium bromide staining. Speed and 
      automation are important considerations for HLA typing since the number of 
      possible alleles for each HLA type is substantial. A homogeneous HLA-SSP typing 
      method may be especially useful for clinical labs doing large numbers of samples 
      and for the eventual automation of HLA DNA typing.
FAU - Okamoto, N
AU  - Okamoto N
AD  - Biomedical Research Center, Olympus America Inc., East Setauket, New York 11733.
FAU - Lee, A
AU  - Lee A
FAU - Kano, T
AU  - Kano T
FAU - Lee, T D
AU  - Lee TD
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 0 (DNA Primers)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (HLA-DR Antigens)
RN  - 0 (Viral Proteins)
RN  - 9007-49-2 (DNA)
RN  - EC 3.1.- (EXO1 protein, human)
RN  - EC 3.1.- (Exodeoxyribonucleases)
RN  - EC 3.1.11.1 (exodeoxyribonuclease I)
RN  - EC 3.1.11.3 (exo protein, Bacteriophage lambda)
RN  - EC 6.5.1.- (DNA Repair Enzymes)
SB  - IM
MH  - Alleles
MH  - Base Sequence
MH  - DNA/blood/isolation & purification
MH  - DNA Primers
MH  - DNA Repair Enzymes
MH  - Exodeoxyribonucleases
MH  - Fluorescent Dyes
MH  - HLA-DR Antigens/*blood/genetics
MH  - Histocompatibility Testing/*methods
MH  - Humans
MH  - Leukocytes/*immunology
MH  - Molecular Sequence Data
MH  - Polymerase Chain Reaction/*methods
MH  - Spectrometry, Fluorescence/methods
MH  - Templates, Genetic
MH  - Viral Proteins
EDAT- 1994/09/01 00:00
MHDA- 1994/09/01 00:01
CRDT- 1994/09/01 00:00
PHST- 1994/09/01 00:00 [pubmed]
PHST- 1994/09/01 00:01 [medline]
PHST- 1994/09/01 00:00 [entrez]
AID - S0003-2697(84)71423-0 [pii]
AID - 10.1006/abio.1994.1423 [doi]
PST - ppublish
SO  - Anal Biochem. 1994 Sep;221(2):340-7. doi: 10.1006/abio.1994.1423.