title: [Development of a method for the separation of HLA-A, -B and -C haploid using biotinylated probe and streptavidin magnetic beads].
abstract: OBJECTIVE
  To develop a method for separating the human leukocyte antigen (HLA)-A, -B and -C haploid using biotinylated probes and streptavidin magnetic beads in order to solve ambiguous HLA genotyping results.
  
  
  METHODS
  Based on sequence information of HLA alleles from the IMGT/HLA database, the 5-biotinylated probes were designed. The probe was mixed and extended with corresponding genomic DNA, and incubated with streptavidin magnetic beads, which could form a streptavidin magnetic beads-biotin-probe DNA complex. The unique DNA haploid binding to corresponding probe was isolated after washes and elution. The separated haploid genomic DNA was used as template for HLA-A, -B and -C loci amplification and sequencing analysis.
  
  
  RESULTS
  Among the 12 HLA-A probes, 19 HLA-B probes and 13 HLA-C probes, DNA sequencing has confirmed that 9 HLA-A probes, 9 HLA-B probes and 5 HLA-C probes could successfully separate the haploid from genomic DNA samples.
  
  
  CONCLUSIONS
  The developed method for HLA-A, -B and -C haploid separation is reliable, which can solve certain ambiguity and improve the accuracy of HLA genotyping.
authors.affiliation: Blood Center of Zhejiang Province, Key Laboratory of Blood Safety Research, Ministry of Health, Hangzhou, Zhejiang 310006, P.R. China. Email: zfm00@hotmail.com.
authors.forename: Sudan
authors.initials: S
authors.lastname: Tao
authors.forename: Yanmin
authors.initials: Y
authors.lastname: He
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Ji
authors.initials: J
authors.lastname: He
authors.forename: Famin
authors.initials: F
authors.lastname: Zhu
authors.forename: Hangjun
authors.initials: H
authors.lastname: Lyu
mesh_terms: D005838:Genotype; D015234:HLA-A Antigens / Q000235:genetics*; D015235:HLA-B Antigens / Q000235:genetics*; D015236:HLA-C Antigens / Q000235:genetics*; D006238:Haploidy; D006801:Humans; D015336:Molecular Probe Techniques / Q000295:instrumentation; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D019809:Streptavidin / Q000737:chemistry
chemical_list: D015234:HLA-A Antigens; D015235:HLA-B Antigens; D015236:HLA-C Antigens; D019809:Streptavidin