Linked Life Data

14969771

Source:http://linkedlifedata.com/resource/pubmed/id/14969771

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2004-2-18
pubmed:abstractText
Sequencing of alleles of the highly polymorphic, multiple loci HLA-DRB gene family was performed by pyrosequencing using purified DNA from the 11(th) International Histocompatibility Workshop human lymphoblastiod cell lines as well as genomic DNA isolated from blood samples obtained from healthy adult volunteers. Genomic DNA was prepared from donors whose blood had been stored either frozen or as dried blood spots. Pyrosequence-based typing was optimized for identifying alleles of the HLA-DRB1, -3, -4, and -5 genes. The procedure should be applicable to other HLA loci including the class I genes HLA-A and -B that, along with HLA-DRB, are crucial for histocompatibility matching of tissue antigens during transplantation. Computer simulation of pyrosequencing data suggest that alleles of HLA-DRB1, -3, -4, and -5 were readily identifiable by pyrosequencing as were their heterozygous allelic combinations. Pyrosequencing primers were designed to specifically sequence HLA loci of interest even in a background of other amplified, closely related sequences such as alleles of the pseudogene HLA-DRB6, -7, -8, and -9. Polymorphic residues of HLA-DRB genes were identified within each pyrosequencing reaction, obtained by 50 to 70 nucleotide read lengths. Heterozygous allelic combinations of HLA genes were analyzed and compared successfully to genotyping of alleles by sequence-specific oligonucleotide probe hybridization as well as allele specific polymerase chain reaction protocols. Pyrosequence-based typing is compatible with genotyping of allelic combinations expected from heterozygous individuals, resulting in nucleotide resolution of the highly polymorphic HLA system. Using a single pyrosequence instrument, complete typing of HLA-DRB genes can be performed daily on hundreds of individuals for high resolution histocompatibility genotyping studies.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0198-8859
pubmed:author
pubmed:issnType
Print
pubmed:volume
65
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
163-74
pubmed:dateRevised
2011-11-17
pubmed:meshHeading
pubmed-meshheading:14969771-Base Sequence, pubmed-meshheading:14969771-Cell Line, pubmed-meshheading:14969771-Cloning, Molecular, pubmed-meshheading:14969771-DNA, pubmed-meshheading:14969771-DNA Primers, pubmed-meshheading:14969771-Genotype, pubmed-meshheading:14969771-HLA-DR Antigens, pubmed-meshheading:14969771-HLA-DR beta-Chains, pubmed-meshheading:14969771-HLA-DRB1 Chains, pubmed-meshheading:14969771-HLA-DRB3 Chains, pubmed-meshheading:14969771-HLA-DRB4 Chains, pubmed-meshheading:14969771-HLA-DRB5 Chains, pubmed-meshheading:14969771-Heterozygote, pubmed-meshheading:14969771-Histocompatibility Antigens Class II, pubmed-meshheading:14969771-Humans, pubmed-meshheading:14969771-Leukocytes, Mononuclear, pubmed-meshheading:14969771-Linear Models, pubmed-meshheading:14969771-Molecular Sequence Data, pubmed-meshheading:14969771-Polymerase Chain Reaction, pubmed-meshheading:14969771-Polymorphism, Genetic, pubmed-meshheading:14969771-Sequence Analysis, DNA, pubmed-meshheading:14969771-Sequence Homology, Nucleic Acid
pubmed:year
2004
pubmed:articleTitle
HLA class II DRB high resolution genotyping by pyrosequencing: comparison of group specific PCR and pyrosequencing primers.
pubmed:affiliation
Department of Pediatrics, Division of Immunogenetics, Rangos Research Center, Children's Hospital of Pittsburgh, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't