High throughput HLA sequence-based typing (SBT) utilizing the ABI Prism 3700 DNA Analyzer.

摘要: Aims and background The genetic complexity of the human major histocompatibility complex (MHC) has required development various molecular typing methods. purpose this paper is to compare results two these methods: sequenced based (SBT) polymerase chain reaction (PCR) using sequence specific primers (PCR-SSP). Methods SBT method described utilizes an ABI Prism 3700 DNA Analyzer, which been designed fro high throughput production data through highly automated operation with significant walk-away time. Analyzer a 96-capillary electrophoresis instrument capability running four 96-well plates black back in sixteen-hour period. Potentially, from machine can produce Class I sequences for A or B loci 64 samples time frame. encompassed exons 2, 3, 4 forward reverse orientation reactions PE Biosystems HLA-A HLA-B Sequenced Based Typing Kits (PE Applied Biopsystems/Perkin-Elmer, Foster City, CA, USA). Most methods previously employed only gather 2 3 distinguishes most polymorphism necessary identify majority alleles HLA region. However, effort discern numerous null region, exon also included. PCR-SSP utilized consists one 96 well tray, 95 primer mixes negative control, per sample intermediate/high resolution HLA-A, typing. Results Data capillary run on 16 loci, was compared derived sixteen typings. 75% tested achieved higher by method. Discussion ability provide allele level have functional implications bone marrow transplant community vaccine studies.