PMID- 20650293
OWN - NLM
STAT- MEDLINE
DCOM- 20110405
LR  - 20211020
IS  - 1879-1166 (Electronic)
IS  - 0198-8859 (Print)
IS  - 0198-8859 (Linking)
VI  - 71
IP  - 10
DP  - 2010 Oct
TI  - A novel single cDNA amplicon pyrosequencing method for high-throughput, 
      cost-effective sequence-based HLA class I genotyping.
PG  - 1011-7
LID - 10.1016/j.humimm.2010.07.012 [doi]
AB  - Human leukocyte antigen (HLA) genotype influences the immune response to 
      pathogens and transplanted tissues; accurate HLA genotyping is critical for 
      clinical and research applications. Sequence-based HLA typing is limited by the 
      cost of Sanger sequencing genomic DNA (gDNA) and resolving cis/trans ambiguities, 
      hindering both studies correlating high-resolution genotype with clinical 
      outcomes, and population-specific allele frequency surveys. We present an assay 
      for sequence-based HLA genotyping by titanium read length clonal Roche/454 
      pyrosequencing of a single, universally diagnostic polymerase chain reaction 
      (PCR) amplicon from HLA class I cDNA that captures most of exons 2, 3, and 4 used 
      for traditional sequence-based typing. The amplicon is predicted to unambiguously 
      resolve 85% of known alleles. A panel of 48 previously HLA-typed samples was 
      assayed with this method, demonstrating 100% non-null allele typing concordance. 
      We show that this technique can multiplex at least 768 patients per sequencing 
      run with multiplex identifier sequence bar-coding. Unprecedented typing 
      throughput results from a novel single cDNA-PCR amplicon strategy requiring only 
      1 PCR amplification per sample. This method dramatically reduces cost for 
      genotyping of large cohorts.
CI  - 2010 American Society for Histocompatibility and Immunogenetics. Published by 
      Elsevier Inc. All rights reserved.
FAU - Lank, Simon M
AU  - Lank SM
AD  - Wisconsin National Primate Research Center, University of Wisconsin-Madison, 
      Madison, Wisconsin, USA.
FAU - Wiseman, Roger W
AU  - Wiseman RW
FAU - Dudley, Dawn M
AU  - Dudley DM
FAU - O'Connor, David H
AU  - O'Connor DH
LA  - eng
GR  - P51 RR000167/RR/NCRR NIH HHS/United States
GR  - P51 RR000167-51/RR/NCRR NIH HHS/United States
GR  - P51 RR000167-50/RR/NCRR NIH HHS/United States
GR  - P51 RR000167-49/RR/NCRR NIH HHS/United States
GR  - P51 OD011106/OD/NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20100730
PL  - United States
TA  - Hum Immunol
JT  - Human immunology
JID - 8010936
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Complementary)
SB  - IM
MH  - Alleles
MH  - Cost-Benefit Analysis
MH  - DNA Primers
MH  - DNA, Complementary/*analysis
MH  - Genes, MHC Class I/*genetics
MH  - High-Throughput Nucleotide Sequencing/methods/trends
MH  - *Histocompatibility Testing/economics
MH  - Humans
MH  - *Nucleic Acid Amplification Techniques/economics
MH  - Sequence Analysis, DNA
PMC - PMC3191540
MID - NIHMS326573
EDAT- 2010/07/24 06:00
MHDA- 2011/04/06 06:00
PMCR- 2011/10/12
CRDT- 2010/07/24 06:00
PHST- 2010/05/26 00:00 [received]
PHST- 2010/07/12 00:00 [revised]
PHST- 2010/07/30 00:00 [accepted]
PHST- 2010/07/24 06:00 [entrez]
PHST- 2010/07/24 06:00 [pubmed]
PHST- 2011/04/06 06:00 [medline]
PHST- 2011/10/12 00:00 [pmc-release]
AID - S0198-8859(10)00447-7 [pii]
AID - 10.1016/j.humimm.2010.07.012 [doi]
PST - ppublish
SO  - Hum Immunol. 2010 Oct;71(10):1011-7. doi: 10.1016/j.humimm.2010.07.012. Epub 2010 
      Jul 30.