title: HLA Typing for the Next Generation.
abstract: Allele-level resolution data at primary HLA typing is the ideal for most histocompatibility testing laboratories. Many high-throughput molecular HLA typing approaches are unable to determine the phase of observed DNA sequence polymorphisms, leading to ambiguous results. The use of higher resolution methods is often restricted due to cost and time limitations. Here we report on the feasibility of using Pacific Biosciences' Single Molecule Real-Time (SMRT) DNA sequencing technology for high-resolution and high-throughput HLA typing. Seven DNA samples were typed for HLA-A, -B and -C. The results showed that SMRT DNA sequencing technology was able to generate sequences that spanned entire HLA Class I genes that allowed for accurate allele calling. Eight novel genomic HLA class I sequences were identified, four were novel alleles, three were confirmed as genomic sequence extensions and one corrected an existing genomic reference sequence. This method has the potential to revolutionize the field of HLA typing. The clinical impact of achieving this level of resolution HLA typing data is likely to considerable, particularly in applications such as organ and blood stem cell transplantation where matching donors and recipients for their HLA is of utmost importance.
authors.affiliation: Anthony Nolan Research Institute, Royal Free Hospital, London, United Kingdom; UCL Cancer Institute, Royal Free Campus, London, United Kingdom.
authors.forename: Neema P
authors.initials: NP
authors.lastname: Mayor
authors.affiliation: Anthony Nolan Research Institute, Royal Free Hospital, London, United Kingdom; UCL Cancer Institute, Royal Free Campus, London, United Kingdom.
authors.forename: James
authors.initials: J
authors.lastname: Robinson
authors.affiliation: Anthony Nolan Research Institute, Royal Free Hospital, London, United Kingdom.
authors.forename: Alasdair J M
authors.initials: AJ
authors.lastname: McWhinnie
authors.affiliation: Pacific Biosciences, Menlo Park, California, United States of America.
authors.forename: Swati
authors.initials: S
authors.lastname: Ranade
authors.affiliation: Pacific Biosciences, Menlo Park, California, United States of America.
authors.forename: Kevin
authors.initials: K
authors.lastname: Eng
authors.affiliation: Anthony Nolan Research Institute, Royal Free Hospital, London, United Kingdom.
authors.forename: William
authors.initials: W
authors.lastname: Midwinter
authors.affiliation: Anthony Nolan Research Institute, Royal Free Hospital, London, United Kingdom.
authors.forename: Will P
authors.initials: WP
authors.lastname: Bultitude
authors.affiliation: Pacific Biosciences, Menlo Park, California, United States of America.
authors.forename: Chen-Shan
authors.initials: CS
authors.lastname: Chin
authors.affiliation: Pacific Biosciences, Menlo Park, California, United States of America.
authors.forename: Brett
authors.initials: B
authors.lastname: Bowman
authors.affiliation: Pacific Biosciences, Menlo Park, California, United States of America.
authors.forename: Patrick
authors.initials: P
authors.lastname: Marks
authors.affiliation: Anthony Nolan Research Institute, Royal Free Hospital, London, United Kingdom.
authors.forename: Henny
authors.initials: H
authors.lastname: Braund
authors.affiliation: Anthony Nolan Research Institute, Royal Free Hospital, London, United Kingdom; UCL Cancer Institute, Royal Free Campus, London, United Kingdom.
authors.forename: J Alejandro
authors.initials: JA
authors.lastname: Madrigal
authors.affiliation: Anthony Nolan Research Institute, Royal Free Hospital, London, United Kingdom.
authors.forename: Katy
authors.initials: K
authors.lastname: Latham
authors.affiliation: Anthony Nolan Research Institute, Royal Free Hospital, London, United Kingdom; UCL Cancer Institute, Royal Free Campus, London, United Kingdom.
authors.forename: Steven G E
authors.initials: SG
authors.lastname: Marsh
mesh_terms: D000483:Alleles; D023281:Genomics; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D017422:Sequence Analysis, DNA / Q000379:methods*