PMID- 17054408
OWN - NLM
STAT- MEDLINE
DCOM- 20061201
LR  - 20061023
IS  - 1462-2416 (Print)
IS  - 1462-2416 (Linking)
VI  - 7
IP  - 7
DP  - 2006 Oct
TI  - Development and clinical evaluation of oligonucleotide microarray for HLA-AB 
      genotyping.
PG  - 973-85
AB  - BACKGROUND: The antigens encoded by human leukocyte antigen (HLA) genes are 
      primary antigens in immunological response of transplantation, and genotypes of 
      HLA-A, B and DRB1 must be determined on donors and recipients before the 
      transplantation is carried out. In this study, oligonucleotide microarray for 
      HLA-AB genotyping was prepared and evaluated. METHODS: Oligonucleotide probes 
      were designed based on the sequence of the different genotypes of HLA-A and B and 
      were fixed on silylated glass slides to form a microarray. Fluorescence-labeled 
      target DNA was obtained by asymmetric polymerase chain reaction amplification of 
      exon 2 and exon 3 of HLA-A and -B genes and hybridized to the microarray. The 
      hybridized microarray was subsequently scanned and the result was analyzed by 
      software in order to determine the genotypes of the tested sample. RESULTS: The 
      sensitivity of the microarray for HLA-AB genotyping was 10 ng/microl, with a 
      coincidence rate of 100% with international reference, and a combined variation 
      value of detected signal below 15%. Analysis of genotyping of 1295 cases of 
      clinical samples showed that the general coincidence rate between the microarray 
      method and conventional method (polymerase chain reaction-sequence-specific 
      primer and flow cytometry reverse polymerase chain reaction sequence-specific 
      oligonucleotide) was HLA-A: 99.61% and HLA-B: 98.30%, respectively. A total of 
      five out of seven samples that had conflicting results of genotypes were proved 
      to be microarray-assay reliable by DNA sequencing, suggesting a higher accuracy 
      of the microarray method. CONCLUSION: The microarray for HLA-AB genotyping is 
      satisfactory for clinical use in HLA-AB genotyping with its good specificity, 
      sensitivity and reproducibility.
FAU - Zhang, Fan
AU  - Zhang F
AD  - Beijing Institute of Radiation Medicine, Beijing 100850, China.
FAU - Hu, Shouwang
AU  - Hu S
FAU - Huang, Jian
AU  - Huang J
FAU - Wang, Hui
AU  - Wang H
FAU - Wen, Zheng
AU  - Wen Z
FAU - Yongyao, Geng
AU  - Yongyao G
FAU - Wang, Shengqi
AU  - Wang S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Pharmacogenomics
JT  - Pharmacogenomics
JID - 100897350
RN  - 0 (DNA Probes)
RN  - 0 (HLA-A Antigens)
RN  - 0 (HLA-B Antigens)
RN  - 0 (Oligonucleotides)
SB  - IM
MH  - DNA Probes
MH  - Double-Blind Method
MH  - Exons/genetics/immunology
MH  - Flow Cytometry
MH  - Genotype
MH  - HLA-A Antigens/*immunology
MH  - HLA-B Antigens/immunology
MH  - Humans
MH  - Nucleic Acid Hybridization
MH  - *Oligonucleotide Array Sequence Analysis
MH  - Oligonucleotides/*genetics
MH  - Reproducibility of Results
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Software
EDAT- 2006/10/24 09:00
MHDA- 2006/12/09 09:00
CRDT- 2006/10/24 09:00
PHST- 2006/10/24 09:00 [pubmed]
PHST- 2006/12/09 09:00 [medline]
PHST- 2006/10/24 09:00 [entrez]
AID - 10.2217/14622416.7.7.973 [doi]
PST - ppublish
SO  - Pharmacogenomics. 2006 Oct;7(7):973-85. doi: 10.2217/14622416.7.7.973.