NDLI: Super high resolution for single molecule-sequence-based typing of classical HLA loci at the 8-digit level using next generation sequencers.

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Super high resolution for single molecule-sequence-based typing of classical HLA loci at the 8-digit level using next generation sequencers.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Shiina, T. Suzuki, S. Ozaki, Y. Taira, H. Kikkawa, E. Shigenari, A. Oka, A. Umemura, T. Joshita, S. Takahashi, O. Hayashi, Y. Paumen, M. Katsuyama, Y. Mitsunaga, S. Ota, M. Kulski, J. K. Inoko, H.
Description	Country affiliation: Japan Author Affiliation: Shiina T ( Division of Basic Medical Science and Molecular Medicine, Department of Molecular Life Science, Tokai University School of Medicine, Isehara, Kanagawa, Japan.)
Abstract	Current human leukocyte antigen (HLA) DNA typing methods such as the sequence-based typing (SBT) and sequence-specific oligonucleotide (SSO) methods generally yield ambiguous typing results because of oligonucleotide probe design limitations or phase ambiguity for HLA allele assignment. Here we describe the development and application of the super high-resolution single-molecule sequence-based typing (SS-SBT) of HLA loci at the 8-digit level using next generation sequencing (NGS). NGS which can determine an HLA allele sequence derived from a single DNA molecule is expected to solve the phase ambiguity problem. Eight classical HLA loci-specific polymerase chain reaction (PCR) primers were designed to amplify the entire gene sequences from the enhancer-promoter region to the 3' untranslated region. Phase ambiguities of HLA-A, -B, -C, -DRB1 and -DQB1 were completely resolved and unequivocally assigned without ambiguity to single HLA alleles. Therefore, the SS-SBT method described here is a superior and effective HLA DNA typing method to efficiently detect new HLA alleles and null alleles without ambiguity.
File Format	HTM / HTML
ISSN	20592302
Issue Number	4
Volume Number	80
e-ISSN	20592310
Journal	Tissue Antigens
Language	English
Publisher	Wiley-Blackwell
Publisher Date	2012-10-01
Publisher Place	Great Britain (UK)
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Genetics Discipline Immunology Genetic Loci Hla Antigens Analysis High-throughput Nucleotide Sequencing Methods Multilocus Sequence Typing 3' Untranslated Regions Alleles Dna Primers Genetics Instrumentation Humans Polymerase Chain Reaction Promoter Regions, Genetic Sequence Analysis, Dna Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Genetics Immunology and Allergy Immunology

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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