PMID- 11476908
OWN - NLM
STAT- MEDLINE
DCOM- 20011011
LR  - 20190906
IS  - 0198-8859 (Print)
IS  - 0198-8859 (Linking)
VI  - 62
IP  - 8
DP  - 2001 Aug
TI  - Molecular typing of HLA-A, -B, and DRB using a high throughput micro array 
      format.
PG  - 850-7
AB  - The goal of this study was to develop a DNA micro array procedure for molecular 
      human leukocyte antigen (HLA) typing of a large number of samples. DNA was 
      isolated from peripheral blood samples and polymerase chain reaction (PCR) 
      amplified for HLA-A, -B, and -DRB. Amplified DNA samples were spotted on 
      silane-treated glass slides using a micro array spotter. The spotter was capable 
      of spotting multiple slides with up to 9216 samples per slide or 2304 samples in 
      quadruplicate. The allele specific oligo nucleotide probes for HLA-A, -B, and 
      -DRB were labeled with the fluorescent dye Cy3, while a control probe, to 
      quantitate the total amount of PCR product in a sample, was labeled with Cy5. 
      Each slide was hybridized with a mixture of an allele specific Cy3 probe plus the 
      control Cy5 probe. Following hybridization and wash, the amount of probe 
      hybridizing to each DNA sample on the slide was measured with a micro array 
      scanner. A computer program was used for image analysis, to calculate the average 
      Cy3/Cy5 ratios and to identify the positive and negative samples. In turn, this 
      information was used to determine the HLA phenotype of each sample. There was 
      very good concordance between the results obtained for all three loci using 
      Cy-labeled probes as compared with those previously obtained by chemiluminescent 
      detection of alkaline phosphatase labeled probes. This methodology has the 
      potential of greatly simplifying HLA molecular typing of large number of samples.
FAU - Balazs, I
AU  - Balazs I
AD  - Lifecodes Corporation, Stamford, CT, USA.
FAU - Beekman, J
AU  - Beekman J
FAU - Neuweiler, J
AU  - Neuweiler J
FAU - Liu, H
AU  - Liu H
FAU - Watson, E
AU  - Watson E
FAU - Ray, B
AU  - Ray B
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PL  - United States
TA  - Hum Immunol
JT  - Human immunology
JID - 8010936
RN  - 0 (Carbocyanines)
RN  - 0 (HLA-A Antigens)
RN  - 0 (HLA-B Antigens)
RN  - 0 (HLA-DR Antigens)
RN  - 0 (cyanine dye 3)
RN  - 0 (cyanine dye 5)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Alleles
MH  - Carbocyanines
MH  - DNA/analysis
MH  - Electronic Data Processing
MH  - HLA-A Antigens/genetics/*immunology
MH  - HLA-B Antigens/genetics/*immunology
MH  - HLA-DR Antigens/genetics/*immunology
MH  - Histocompatibility Testing/*methods
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
EDAT- 2001/07/31 10:00
MHDA- 2001/10/12 10:01
CRDT- 2001/07/31 10:00
PHST- 2001/07/31 10:00 [pubmed]
PHST- 2001/10/12 10:01 [medline]
PHST- 2001/07/31 10:00 [entrez]
AID - S0198-8859(01)00274-9 [pii]
AID - 10.1016/s0198-8859(01)00274-9 [doi]
PST - ppublish
SO  - Hum Immunol. 2001 Aug;62(8):850-7. doi: 10.1016/s0198-8859(01)00274-9.